Targeted antimicrobial moieties

ABSTRACT

This invention provides novel targeted antimicrobial compositions. In various embodiments chimeric moieties are provided comprising an antimicrobial peptide attached to a peptide targeting moiety that binds a bacterial strain or species.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.61/150,287, filed Feb. 5, 2009; and to U.S. Provisional Application No.61/224,825, filed Jul. 10, 2009, the disclosures of which areincorporated herein by reference in their entirety for all purposes.

STATEMENT OF GOVERNMENTAL SUPPORT

[Not Applicable]

FIELD OF THE INVENTION

The present invention relates to novel targeting peptides, novelantimicrobial peptides, chimeric moieties comprising novel targetingand/or novel antimicrobial peptides and uses thereof.

BACKGROUND OF THE INVENTION

Antibiotic research at the industrial level was originally focused onthe identification of refined variants of already existing drugs. Thisresulted example, in the development of antibiotics such as newerpenicillins, cephalosporins, macrolides, and fluoroquinolones.

However, resistance to old and newer antibiotics among bacterialpathogens is evolving rapidly, as exemplified by extended beta-lactamase(ESBL) and quinolone resistant gram-negatives, multi-resistantgonococci, methicillin resistant Staphylococcus aureus (MRSA),vancomycin resistant enterococci (VRE), penicillin non-susceptiblepneumococci (PNSP) and macrolide resistant pneumococci and streptococci(see, e.g., Panlilo et al. (1992) Infect. Control Hosp. EpidemioL., 13:582-586; Morris et al. (1995) Ann Intern Med., 123: 250-259, and thelike). An overuse, or improper use, of antibiotics is believed to be ofgreat importance for triggering and spread of drug resistant bacteria.Microbes have, in many cases, adapted and are resistant to antibioticsdue to constant exposure and improper use of the drugs.

Drug resistant pathogens represent a major economic burden forhealth-care systems. For example, postoperative and other nosocomialinfections will prolong the need for hospital care and increaseantibiotic drug expenses. It is estimated that the annual cost oftreating drug resistant infections in the United States is approximately$5 billion.

SUMMARY OF THE INVENTION

In certain embodiments, novel targeting moieties (e.g., peptides) thatspecifically/preferentially bind to microorganisms (e.g., certainbacteria, yeasts, fungi, molds, viruses, algae, protozoa, and the like)are provided. The targeting moieties can be attached to effectors (e.g.,detectable labels, drugs, antimicrobial peptides, etc.) to form chimericconstructs for specifically/preferentially delivering the effector toand/or into the target organism. In certain embodiments novelantimicrobial peptides that can be used to inhibit (e.g., kill and/orinhibit growth and/or proliferation) of certain microorganisms (e.g.,certain bacteria, yeasts, fungi, molds, viruses, algae, protozoa, andthe like) are provided. Any targeting moiety disclosed herein can beattached to any one or more effector disclosed herein. Any targetingmoiety disclosed herein can be attached to any one or more antimicrobialpeptide disclosed herein.

In certain embodiments chimeric moieties are provided where the chimericmoieties comprise an effector attached to a peptide targeting moietycomprising or consisting of the amino acid sequence of a peptide foundin Table 2 and/or Table 15. In certain embodiments the targeting peptidecomprises or consists of amino acid or retro or inverso or retro-inversosequence or beta sequence of a peptide found in Table 2 and/or Table 15.In certain embodiments the effector comprises a moiety selected from thegroup consisting of a detectable label, an antimicrobial peptide, anantibiotic, and a photosensitizer. In certain embodiments the effectorcomprises an antimicrobial peptide comprising the amino acid sequence ofa peptide found in Table 2, and/or Table 8, and/or Table 9, and/or Table10. In certain embodiments the effector comprises an antibiotic found inTable 7. In certain embodiments the effector comprises aphotosensitizer. In certain embodiments the photosensitizer is selectedfrom the group consisting of a porphyrinic macrocycle, a porphyrin, achlorine, a crown ether, an acridine, an azine, a phthalocyanine, acyanine, a psoralen, and a perylenequinonoid. In certain embodiments thephotosensitizing agent is an agent shown in any of FIGS. 1-11.

Also provided is a chimeric construct comprising a targeting moietyattached to an antimicrobial peptide where the antimicrobial peptidecomprises or consists of the amino acid or retro or inverso orretro-inverso sequence of a peptide found in Table 2. In certainembodiments the targeting moiety is a peptide that comprises or consistsof the amino acid or retro or inverso or retro-inverso or beta sequenceof a peptide found in Table 2, and/or Table 3, and/or Table 4, and/orTable 6, and/or Table 15. In certain embodiments, the targeting moietycomprises an antibody (e.g., an antibody identified in Table 5). Incertain embodiments the targeting moiety is chemically conjugated to theeffector directly or via a linker. In certain embodiments the targetingmoiety is chemically conjugated to the effector via a linker comprisinga polyethylene glycol (PEG). In certain embodiments the targeting moietyis chemically conjugated to the effector via a non-peptide linker foundin Table 11. In certain embodiments the where the targeting moiety islinked to the effector via a peptide linkage. In certain embodiments thechimeric construct is a fusion protein. In certain embodiments thelinker is a peptide linker found in Table 11. In certain embodiments thechimeric moiety is functionalized with a polymer (e.g., polyethyleneglycol, a cellulose, a modified cellulose, etc.) to increase serumhalflife.

Also provided are pharmaceutical compositions comprising the chimericconstruct(s)/chimeric moieties described herein in a pharmaceuticallyacceptable carrier. In certain embodiments the composition is formulatedas a unit dosage formulation. In certain embodiments the composition isformulated for administration by a modality selected from the groupconsisting of intraperitoneal administration, topical administration,oral administration, inhalation administration, transdermaladministration, subdermal depot administration, and rectaladministration.

Also provided is an antimicrobial composition comprising an isolatedantimicrobial moiety comprising or consisting of the amino acid sequenceof a peptide found in Table 2. In certain embodiments the peptidecomprises or consists of the amino acid or retro or inverso orretro-inverso sequence or beta sequence of a peptide found in Table 2.In certain embodiments the peptide is a peptide selected from the groupconsisting of a peptide consisting of the amino acid sequence of apeptide found in Table 2 comprising all L residues, a peptide consistingof the amino acid sequence of a peptide found in Table 2 comprising apeptide found in Table comprising all D residues, a peptide comprisingthe inverse of an amino acid sequence found in Table 2, a peptidecomprising the retro-inverso form of a peptide found in Table 2, apeptide found in Table 2 comprising a conservative substitution, and apeptide found in Table 2 comprising a substitution of a naturallyoccurring amino acid with a non-naturally occurring amino acid. Incertain embodiments the peptide comprises no more than 1, 2, 3, 4, 5, 6,7, 8, 9, or 10 conservative substitutions.

Also provided is a composition comprising an isolated targeting moietycomprising or consisting of the amino acid sequence of a peptide foundin Table 2 or Table 15. In certain embodiments the peptide comprises orconsists of the amino acid or retro or inverso or retro-inverso sequenceor beta sequence of a peptide found in Table 2 or Table 15. In certainembodiments the peptide is a peptide selected from the group consistingof a peptide consisting of the amino acid sequence of a peptide found inTable 2 or Table 15 comprising all L residues, a peptide consisting ofthe amino acid sequence of a peptide found in Table 2 comprising apeptide found in Table comprising all D residues, a peptide comprisingthe inverse of an amino acid sequence found in Table 2 or Table 15, apeptide comprising the retro-inverso form of a peptide found in Table 2or Table 15, a peptide found in Table 2 or Table 15 comprising aconservative substitution, and a peptide found in Table 2 or Table 15comprising a substitution of a naturally occurring amino acid with anon-naturally occurring amino acid. In certain embodiments the peptidecomprises no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 conservativesubstitutions.

In certain embodiments methods are provided for inhibiting the growthand/or proliferation of a microorganism and/or a biofilm comprising amicroorganism. The methods typically involve contacting themicroorganism or biofilm with a composition comprising an antimicrobialpeptide comprising or consisting of the amino acid sequence of a peptidefound in Table 2 (e.g., the amino acid or retro or inverso orretro-inverso sequence or beta sequence of a peptide found in Table 2);and/or contacting the microorganism or biofilm with a compositioncomprising an antimicrobial moiety attached to a targeting peptidecomprising or consisting of the amino acid sequence of a peptide foundin Table 2 (e.g., the amino acid or retro or inverso or retro-inversosequence or beta sequence of a peptide found in Table 2). In certainembodiments the microorganism or biofilm is a bacterium or a bacterialfilm. In certain embodiments the targeting peptide is chemicallyconjugated to the antimicrobial peptide. In certain embodiments thetargeting peptide is linked directly to the antimicrobial peptide. Incertain embodiments the targeting peptide is linked to the antimicrobialpeptide via a linker comprising a polyethylene glycol. In certainembodiments the targeting peptide is linked to the antimicrobial peptidevia a non-peptide linkage in Table 11. In certain embodiments thetargeting peptide is linked to the antimicrobial peptide via a peptidelinkage. In certain embodiments the targeting peptide linked to theantimicrobial peptide is a fusion protein. In certain embodiments thelinker is a peptide linker in Table 11.

In various embodiments methods are provided for detecting amicroorganism (e.g., bacteria, yeast, protozoan, virus, algae, fungi,etc.) or biofilm comprising the microorganism. The methods typicallyinvolve contacting the microorganism or biofilm with a compositioncomprising a detectable label attached to a targeting peptide comprisingthe amino acid sequence of a peptide comprising or consisting of theamino acid or retro or inverso or retro-inverso sequence of a peptidefound in Table 2; and detecting the detectable label where the quantityand/or location of the detectable label is an indicator of the presenceof the microorganism and/or biofilm film. In certain embodiments themicroorganism or biofilm is a bacterium or a bacterial film. In certainembodiments the detectable label is a label selected from the groupconsisting of a radioactive label, a radio-opaque label, a fluorescentdye, a fluorescent protein, an enzymatic label, a colorimetric label,and a quantum dot.

In certain embodiments compositions are also provided comprising aphotosensitizing agent attached to a targeting peptide where thetargeting peptide comprising or consisting of the amino acid or retro orinverso or retro-inverso sequence of a peptide found in Table 2 or Table15. In certain embodiments the photosensitizing agent is an agentselected from the group consisting of a porphyrinic macrocycle, aporphyrin, a chlorine, a crown ether, an acridine, an azine, aphthalocyanine, a cyanine, a psoralen, and a perylenequinonoid. Incertain embodiments the photosensitizing agent is an agent shown in anyof FIGS. 1-11. In certain embodiments the photosensitizing agent isattached to the targeting peptide by a non-peptide linker. In certainembodiments photosensitizing agent is attached to the targeting peptideby a linker comprising a polyethylene glycol (PEG). In certainembodiments the photosensitizing agent is attached to the targetingpeptide by a non-peptide linker found in Table 11.

In certain embodiments methods are provided for inhibiting the growth orproliferation of a microorganism and/or a biofilm (e.g., a bacteriumand/or a bacterial film), where the methods involve contacting the amicroorganism and/or a biofilm with a composition comprising aphotosensitizing agent attached to a targeting peptide as describedherein. In certain embodiments the method further comprises exposing themicroorganism or biofilm to a light source. In certain embodiments themicroorganism is a microorganism selected from the group consisting of abacterium, a yeast, a fungus, a protozoan, and a virus. In certainembodiments the biofilm comprises a bacterial film.

In certain embodiments chimeric moieties are provided wherein thechimeric moiety comprises multiple targeting moieties attached to eachother. In certain embodiments the targeting moieties are directlyattached to each other. In certain embodiments the targeting moietiesare attached to each other via a peptide linker. In certain embodimentsthe targeting moieties are attached to each other via a non-peptidelinker. In certain embodiments chimeric moieties are provided whereinthe chimeric moiety comprises multiple effectors attached to each other.In certain embodiments the effectors are directly attached to eachother. In certain embodiments the effectors are attached to each othervia a peptide linker. In certain embodiments the effectors are attachedto each other via a non-peptide linker.

In certain embodiments chimeric moieties are provided where the chimericmoiety comprises one or more targeting moieties attached to one or moreeffectors. In certain embodiments the chimeric moiety comprises one ormore of the targeting moieties shown in Table 2, and/or Table 4 and/orTable 6, and/or Table 15 attached to a single effector. In certainembodiments the chimeric moiety comprises one or more effectors attachedto a single targeting moiety. In certain embodiments the chimeric moietycomprises one or more effectors comprising one or more of theantimicrobial peptides shown in Table 2, and/or Table 8, and/or Table 9,and/or Table 10 attached to a single targeting moiety. In certainembodiments the chimeric moiety comprises multiple targeting moietiesattached to multiple effectors.

DEFINITIONS

The term “peptide” as used herein refers to a polymer of amino acidresidues typically ranging in length from 2 to about 50 residues. Incertain embodiments the peptide ranges in length from about 2, 3, 4, 5,7, 9, 10, or 11 residues to about 50, 45, 40, 45, 30, 25, 20, or 15residues. In certain embodiments the peptide ranges in length from about8, 9, 10, 11, or 12 residues to about 15, 20 or 25 residues. Where anamino acid sequence is provided herein, L-, D-, or beta amino acidversions of the sequence are also contemplated as well as retro,inversion, and retro-inversion isoforms. Peptides also include aminoacid polymers in which one or more amino acid residues is an artificialchemical analogue of a corresponding naturally occurring amino acid, aswell as to naturally occurring amino acid polymers. In addition, theterm applies to amino acids joined by a peptide linkage or by other,“modified linkages” (e.g., where the peptide bond is replaced by anα-ester, a β-ester, a thioamide, phosphonamide, carbomate, hydroxylate,and the like (see, e.g., Spatola, (1983) Chem. Biochem. Amino Acids andProteins 7: 267-357), where the amide is replaced with a saturated amine(see, e.g., Skiles et al., U.S. Pat. No. 4,496,542, which isincorporated herein by reference, and Kaltenbronn et al., (1990) Pp.969-970 in Proc. 11th American Peptide Symposium, ESCOM SciencePublishers, The Netherlands, and the like)).

The term “residue” as used herein refers to natural, synthetic, ormodified amino acids. Various amino acid analogues include, but are notlimited to 2-aminoadipic acid, 3-aminoadipic acid, beta-alanine(beta-aminopropionic acid), 2-aminobutyric acid, 4-aminobutyric acid,piperidinic acid, 6-aminocaproic acid, 2-aminoheptanoic acid,2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, 2,4diaminobutyric acid, desmosine, 2,2′-diaminopimelic acid,2,3-diaminopropionic acid, n-ethylglycine, n-ethylasparagine,hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline,isodesmosine, allo-isoleucine, n-methylglycine, sarcosine,n-methylisoleucine, 6-n-methyllysine, n-methylvaline, norvaline,norleucine, ornithine, and the like. These modified amino acids areillustrative and not intended to be limiting.

“β-peptides” comprise of “β amino acids”, which have their amino groupbonded to the β carbon rather than the α-carbon as in the 20 standardbiological amino acids. The only commonly naturally occurring β aminoacid is β-alanine.

Peptoids, or N-substituted glycines, are a specific subclass ofpeptidomimetics. They are closely related to their natural peptidecounterparts, but differ chemically in that their side chains areappended to nitrogen atoms along the molecule's backbone, rather than tothe α-carbons (as they are in natural amino acids).

The terms “conventional” and “natural” as applied to peptides hereinrefer to peptides, constructed only from the naturally-occurring aminoacids: Ala, Cys, Asp, Glu, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val, Trp, and Tyr. A compound of the invention“corresponds” to a natural peptide if it elicits a biological activity(e.g., antimicrobial activity) related to the biological activity and/orspecificity of the naturally occurring peptide. The elicited activitymay be the same as, greater than or less than that of the naturalpeptide. In general, such a peptoid will have an essentiallycorresponding monomer sequence, where a natural amino acid is replacedby an N-substituted glycine derivative, if the N-substituted glycinederivative resembles the original amino acid in hydrophilicity,hydrophobicity, polarity, etc. Thus, for example, the following pairs ofpeptides would be considered “corresponding”:

(SEQ ID NO: 1) Ia. Asp-Arg-Val-Tyr-Ile-His-Pro-Phe (Angiotensin II) and(SEQ ID NO: 2) Ib. Asp-Arg-Val*-Tyr-Ile*-His-Pro-Phe; (SEQ ID NO: 3)IIa. Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg (Bradykinin) and (SEQ ID NO: 4)IIb: Arg-Pro-Pro-Gly-Phe*-Ser*-Pro-Phe*-Arg; (SEQ ID NO: 5) IIIa:Gly-Gly-Phe-Met-Thr-Ser-Glu-Lys-Ser-Gln-Thr- Pro-Leu-Val-Thr(β-Endorphin); and (SEQ ID NO: 6) IIIb:Gly-Gly-Phe*-Met-Ser*-Ser-Glu-Lys*-Ser-Gln- Ser*-Pro-Leu-Val*-Thr.In these examples, “Val*” refers to N-(prop-2-yl)glycine, “Phe*” refersto N-benzylglycine, “Ser*” refers to N-(2-hydroxyethyl)glycine, “Leu*”refers to N-(2-methylprop-1-yl)glycine, and “Ile*” refers toN-(1-methylprop-1-yl)glycine. The correspondence need not be exact: forexample, N-(2-hydroxyethyl)glycine may substitute for Ser, Thr, Cys, andMet; N-(2-methylprop-1-yl)glycine may substitute for Val, Leu, and Ile.Note in IIIa and IIIb above that Ser* is used to substitute for Thr andSer, despite the structural differences: the sidechain in Ser* is onemethylene group longer than that of Ser, and differs from Thr in thesite of hydroxy-substitution. In general, one may use anN-hydroxyalkyl-substituted glycine to substitute for any polar aminoacid, an N-benzyl- or N-aralkyl-substituted glycine to replace anyaromatic amino acid (e.g., Phe, Trp, etc.), an N-alkyl-substitutedglycine such as N-butylglycine to replace any nonpolar amino acid (e.g.,Leu, Val, Ile, etc.), and an N-(aminoalkyl)glycine derivative to replaceany basic polar amino acid (e.g., Lys and Arg).

A “compound antimicrobial peptide” or “compound AMP” refers to aconstruct comprising two or more AMPs joined together. The AMPs can bejoined directly or through a linker. They can be chemically conjugatedor, where joined directly together or through a peptide linker cancomprise a fusion protein.

In certain embodiments, conservative substitutions of the amino acidscomprising any of the sequences described herein are contemplated. Invarious embodiments one, two, three, four, or five different residuesare substituted. The term “conservative substitution” is used to reflectamino acid substitutions that do not substantially alter the activity(e.g., antimicrobial activity and/or specificity) of the molecule.Typically conservative amino acid substitutions involve substitution oneamino acid for another amino acid with similar chemical properties (e.g.charge or hydrophobicity). Certain conservative substitutions include“analog substitutions” where a standard amino acid is replaced by anon-standard (e.g., rare, synthetic, etc) amino acid differing minimallyfrom the parental residue. Amino acid analogs are considered to bederived synthetically from the standard amino acids without sufficientchange to the structure of the parent, are isomers, or are metaboliteprecursors. Examples of such “analog substitutions” include, but are notlimited to, 1) Lys-Orn, 2) Leu-Norleucine, 3) Lys-Lys[TFA], 4)Phe-Phe[Gly], and 5) 6-amino butylglycine-4-amino hexylglycine, wherePhe[gly] refers to phenylglycine (a Phe derivative with a H rather thanCH₃ component in the R group), and Lys[TFA] refers to a Lys where anegatively charged ion (e.g., TFA) is attached to the amine R group.Other conservative substitutions include “functional substitutions”where the general chemistries of the two residues are similar, and canbe sufficient to mimic or partially recover the function of the nativepeptide. Strong functional substitutions include, but are not limitedto 1) Gly/Ala, 2) Arg/Lys, 3) Ser/Tyr/Thr, 4) Leu/Ile/Val, 5) Asp/Glu,6) Gln/Asn, and 7) Phe/Trp/Tyr, while other functional substitutionsinclude, but are not limited to 8) Gly/Ala/Pro, 9) Tyr/His, 10)Arg/Lys/His, 11) Ser/Thr/Cys, 12) Leu/Ile/Val/Met, and 13) Met/Lys(special case under hydrophobic conditions). Various “broad conservativesubstations” include substitutions where amino acids replace other aminoacids from the same biochemical or biophysical grouping. This issimilarity at a basic level and stems from efforts to classify theoriginal 20 natural amino acids. Such substitutions include 1) nonpolarside chains: Gly/Ala/Val/Leu/Ile/Met/Pro/Phe/Trp, and/or 2) unchargedpolar side chains Ser/Thr/Asn/Gln/Tyr/Cys. In certain embodimentsbroad-level substitutions can also occur as paired substitutions. Forexample, any hydrophilic neutral pair [Ser, Thr, Gln, Asn, Tyr,Cys]+[Ser, Thr, Gln, Asn, Tyr, Cys] can may be replaced by acharge-neutral charged pair [Arg, Lys, His]+[Asp, Glu]. The followingsix groups each contain amino acids that are typical conservativesubstitutions for one another: 1) Alanine (A), Serine (S), Threonine(T); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N),Glutamine (Q); 4) Arginine (R), Lysine (K), Histidine (H); 5) Isoleucine(I), Leucine (L), Methionine (M), Valine (V); and 6) Phenylalanine (F),Tyrosine (Y), Tryptophan (W).

In certain embodiments, targeting peptides, antimicrobial peptides,and/or STAMPs compromising at least 80%, preferably at least 85% or 90%,and more preferably at least 95% or 98% sequence identity with any ofthe sequences described herein are also contemplated. The terms“identical” or percent “identity,” refer to two or more sequences thatare the same or have a specified percentage of amino acid residues thatare the same, when compared and aligned for maximum correspondence, asmeasured using one of the following sequence comparison algorithms or byvisual inspection. With respect to the peptides of this inventionsequence identity is determined over the full length of the peptide. Forsequence comparison, typically one sequence acts as a referencesequence, to which test sequences are compared. When using a sequencecomparison algorithm, test and reference sequences are input into acomputer, subsequence coordinates are designated, if necessary, andsequence algorithm program parameters are designated. The sequencecomparison algorithm then calculates the percent sequence identity forthe test sequence(s) relative to the reference sequence, based on thedesignated program parameters. Optimal alignment of sequences forcomparison can be conducted, e.g., by the local homology algorithm ofSmith & Waterman (1981) Adv. Appl. Math. 2: 482, by the homologyalignment algorithm of Needleman & Wunsch (1970) J. Mol. Biol. 48: 443,by the search for similarity method of Pearson & Lipman (1988) Proc.Natl. Acad. Sci., USA, 85: 2444, by computerized implementations ofthese algorithms (GAP, BESTFIT, FASTA, and TFASTA in the WisconsinGenetics Software Package, Genetics Computer Group, 575 Science Dr.,Madison, Wis.), or by visual inspection.

The term “specificity” when used with respect to the antimicrobialactivity of a peptide indicates that the peptide preferentially inhibitsgrowth and/or proliferation and/or kills a particular microbial speciesas compared to other related and/or unrelated microbes. In certainembodiments the preferential inhibition or killing is at least 10%greater (e.g., LD₅₀ is 10% lower), preferably at least 20%, 30%, 40%, or50%, more preferably at least 2-fold, at least 5-fold, or at least10-fold greater for the target species.

“Treating” or “treatment” of a condition as used herein may refer topreventing the condition, slowing the onset or rate of development ofthe condition, reducing the risk of developing the condition, preventingor delaying the development of symptoms associated with the condition,reducing or ending symptoms associated with the condition, generating acomplete or partial regression of the condition, or some combinationthereof.

The term “consisting essentially of” when used with respect to anantimicrobial peptide (AMP) or AMP motif as described herein, indicatesthat the peptide or peptides encompassed by the library or variants,analogues, or derivatives thereof possess substantially the same orgreater antimicrobial activity and/or specificity as the referencedpeptide. In certain embodiments substantially the same or greaterantimicrobial activity indicates at least 80%, preferably at least 90%,and more preferably at least 95% of the anti microbial activity of thereferenced peptide(s) against a particular bacterial species (e.g., S.mutans).

The term “porphyrinic macrocycle” refers to a porphyrin or porphyrinderivative. Such derivatives include porphyrins with extra ringsortho-fused, or orthoperifused, to the porphyrin nucleus, porphyrinshaving a replacement of one or more carbon atoms of the porphyrin ringby an atom of another element (skeletal replacement), derivatives havinga replacement of a nitrogen atom of the porphyrin ring by an atom ofanother element (skeletal replacement of nitrogen), derivatives havingsubstituents other than hydrogen located at the peripheral (meso-,.beta.-) or core atoms of the porphyrin, derivatives with saturation ofone or more bonds of the porphyrin (hydroporphyrins, e.g., chlorins,bacteriochlorins, isobacteriochlorins, decahydroporphyrins, corphins,pyrrocorphins, etc.), derivatives obtained by coordination of one ormore metals to one or more porphyrin atoms (metalloporphyrins),derivatives having one or more atoms, including pyrrolic andpyrromethenyl units, inserted in the porphyrin ring (expandedporphyrins), derivatives having one or more groups removed from theporphyrin ring (contracted porphyrins, e.g., corrin, corrole) andcombinations of the foregoing derivatives (e.g. phthalocyanines,porphyrazines, naphthalocyanines, subphthalocyanines, and porphyrinisomers). Certain porphyrinic macrocycles comprise at least one5-membered ring.

As used herein, an “antibody” refers to a protein consisting of one ormore polypeptides substantially encoded by immunoglobulin genes orfragments of immunoglobulin genes. The recognized immunoglobulin genesinclude the kappa, lambda, alpha, gamma, delta, epsilon and mu constantregion genes, as well as myriad immunoglobulin variable region genes.Light chains are classified as either kappa or lambda. Heavy chains areclassified as gamma, mu, alpha, delta, or epsilon, which in turn definethe immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively.

A typical immunoglobulin (antibody) structural unit is known to comprisea tetramer. Each tetramer is composed of two identical pairs ofpolypeptide chains, each pair having one “light” (about 25 kD) and one“heavy” chain (about 50-70 kD). The N-terminus of each chain defines avariable region of about 100 to 110 or more amino acids primarilyresponsible for antigen recognition. The terms variable light chain(V_(L)) and variable heavy chain (V_(H)) refer to these light and heavychains respectively.

Antibodies exist as intact immunoglobulins or as a number of wellcharacterized fragments produced by digestion with various peptidases.Thus, for example, pepsin digests an antibody below the disulfidelinkages in the hinge region to produce F(ab)′₂, a dimer of Fab whichitself is a light chain joined to V_(H)-C_(H)1 by a disulfide bond. TheF(ab)′₂ may be reduced under mild conditions to break the disulfidelinkage in the hinge region thereby converting the (Fab′)₂ dimer into anFab′ monomer. The Fab′ monomer is essentially an Fab with part of thehinge region (see, Fundamental Immunology, W. E. Paul, ed., Raven Press,N.Y. (1993), for a more detailed description of other antibodyfragments). While various antibody fragments are defined in terms of thedigestion of an intact antibody, one of skill will appreciate that suchFab′ fragments may be synthesized de novo either chemically or byutilizing recombinant DNA methodology. Thus, the term antibody, as usedherein also includes antibody fragments either produced by themodification of whole antibodies or synthesized de novo usingrecombinant DNA methodologies, including, but are not limited to, Fab′₂,IgG, IgM, IgA, scFv, dAb, nanobodies, unibodies, and diabodies.

In certain embodiments antibodies and fragments of the present inventioncan be bispecific. Bispecific antibodies or fragments can be of severalconfigurations. For example, bispecific antibodies may resemble singleantibodies (or antibody fragments) but have two different antigenbinding sites (variable regions). In various embodiments bispecificantibodies can be produced by chemical techniques (Kranz et al. (1981)Proc. Natl. Acad. Sci., USA, 78: 5807), by “polydoma” techniques (see,e.g., U.S. Pat. No. 4,474,893), or by recombinant DNA techniques. Incertain embodiments bispecific antibodies of the present invention canhave binding specificities for at least two different epitopes, at leastone of which is an epitope of a microbial organism. The microbialbinding antibodies and fragments can also be heteroantibodies.Heteroantibodies are two or more antibodies, or antibody bindingfragments (e.g., Fab) linked together, each antibody or fragment havinga different specificity.

The term “STAMP” refers to Specifically Targeted Anti-MicrobialPeptides. An MH-STAMP is a STAMP bearing two or more targeting domains(i.e., a multi-headed STAMP).

In various embodiments the amino acid abbreviations shown in Table 1 areused herein.

TABLE 1 Amino acid abbreviations. Abbreviation Name 3 Letter 1 LetterAlanine Ala A βAlanine (NH₂—CH₂—CH₂—COOH) βAla Arginine Arg R AsparagineAsn N Aspartic Acid Asp D Cysteine Cys C Glutamic Acid Glu E GlutamineGln Q Glycine Gly G Histidine His H Homoserine Hse — Isoleucine Ile ILeucine Leu L Lysine Lys K Methionine Met M Methionine sulfoxide Met (O)— Methionine methylsulfonium Met (S-Me) — Norleucine Nle — PhenylalaninePhe F Proline Pro P Serine Ser S Threonine Thr T Tryptophan Trp WTyrosine Tyr Y Valine Val V episilon-aminocaproic acid Ahx J(NH²—(CH₂)₅—COOH) 4-aminobutanoic acid gAbu (NH₂(CH₂)₃—COOH)tetrahydroisoquinoline-3- O carboxylic acidLys(N(epsilon)-trifluoroacetyl) K[TFA] α-aminoisobutyric acid Aib B

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows some illustrative porphyrins (compounds 92-99) suitable foruse as targeting moieties and/or antimicrobial effectors.

FIG. 2 shows some illustrative porphyrins (compounds 100-118) suitablefor use as targeting moieties and/or antimicrobial effectors.

FIG. 3 shows some illustrative porphyrins (in particularphthalocyanines) (compounds 119-128) suitable for use as targetingmoieties and/or antimicrobial effectors.

FIG. 4 illustrates the structures of two phthalocyanines, MonoastralFast Blue B and Monoastral Fast Blue G suitable for use as targetingmoieties and/or antimicrobial effectors.

FIG. 5 illustrates certain azine photosensitizers suitable for use astargeting moieties and/or antimicrobial effectors in the compositionsand methods described herein.

FIG. 6 shows illustrative cyanine suitable for use as targeting moietiesand/or antimicrobial effectors in the compositions and methods describedherein.

FIG. 7 shows illustrative psoralen (angelicin) photosensitizers suitablefor use as targeting moieties and/or antimicrobial effectors in thecompositions and methods described herein.

FIG. 8 shows illustrative hypericin and the perylenequinonoid pigmentssuitable for use as targeting moieties and/or antimicrobial effectors inthe compositions and methods described herein.

FIG. 9 shows illustrative acridines suitable for use as targetingmoieties and/or antimicrobial effectors in the compositions and methodsdescribed herein.

FIG. 10 illustrates the structure of the acridine Rose Bengal.

FIG. 11 illustrates various crown ethers suitable for use as targetingmoieties and/or antimicrobial effectors in the compositions and methodsdescribed herein.

FIG. 12 schematically shows some illustrative configurations forchimeric constructs described herein. A: Shows a single targeting moietyT1 attached to a single effector E1 by a linker/spacer L. B: Showsmultiple targeting moieties T1, T2, T3 attached directly to each otherand attached by a linker L to a single effector E1. In variousembodiments T1, T2, and T3, can be domains in a fusion protein. C: Showsmultiple targeting moieties T1, T2, T3 attached to each other by linkersL and attached by a linker L to a single effector E1. In variousembodiments T1, T2, and T3, can be domains in a fusion protein. D: Showsa single targeting moiety T1 attached by a linker L to multipleeffectors E1, E2, and E3 joined directly to each other. E: Shows asingle targeting moiety T1 attached by a linker L to multiple effectorsE1, E2, and E3 joined to each other by linkers L. F: Shows multipletargeting moieties joined directly to each other and by a linker L tomultiple effectors joined to each other by linkers L. G: Shows multipletargeting moieties joined to each other by linkers L and by a linker Lto multiple effectors joined to each other by linkers L. In variousembodiments T1, T2, and T3, and/or E1, E2, and E3 can be domains in afusion protein. H: Illustrates a branched configuration where multipletargeting moieties are linked to a single effector. I: Illustrates adual branched configuration where multiple targeting moieties are linkedto multiple effectors. J: Illustrates a branched configuration wheremultiple targeting moieties are linked to multiple effectors where theeffectors are joined to each other in a linear configuration.

FIG. 13 illustrates various MH-STAMPs used in Example 1. The design,sequence, and observed mass (m/z) for M8(KH)-20 (SEQ ID NO:7), BL(KH)-20(SEQ ID NO:8), and M8(BL)-20 (SEQ ID NO:9.

FIGS. 14A and 14B show HPLC and MS spectra of M8(KH)-20. The quality ofthe completed MH-STAMP was analyzed by HPLC (FIG. 14A) and MALDI massspectroscopy (FIG. 14B). At UV absorbance 215 nm (260 and 280 nm arealso plotted), a single major product was detected by HPLC (* retentionvolume 11.04 mL). After fraction collection, the correct mass (m/z) forsingle-charged M8(KH)-20, 4884.91 (marked by *), was observed for thispeak. Y-axis: 14A, mAU miliabsorbance units; 14B, percent intensity.

FIG. 15A-15E show growth inhibitory activity of MH-STAMPs. Monoculturesof S. mutans (FIG. 15A); P. aeruginosa (FIG. 15B); S. epidermidis (FIG.15C); S. aureus (FIG. 15D); or E. coli (FIG. 15E); were treated withpeptides (as indicated in the figure) for 10 min. Agent was then removedand fresh media returned. Culture recovery was measured over time(OD600). Plots represent the average of at least 3 independentexperiments with standard deviations.

FIG. 16 illustrates the selective activity of dual-targeted andsingle-targeted MH-STAMPs in mixed culture. A mixture of P. aeruginosa(Pa), S. mutans (Sm), E. coli (Ec), and S. epidermidis (Se) planktoniccells were mixed with MH-STAMPs (as indicated in the figure) and treated24 h. After incubation, cfu/mL of remaining constituent species werequantitated after plating to selective media. * indicates under 200surviving cfu/mL recovered.

FIG. 17 depicts the results of minimal inhibitory concentration (MIC)assays, conducted against S. mutans and a panel of bacteria, includingtwo oral streptococci, S. sanguinis and S. sobrinus, to gauge Library 1STAMP antimicrobial activity and S. mutans-selectivity.

FIG. 18 shows the results of MIC assays utilizing targeting peptide2_(—)1 conjugated to one of five AMPs, RWRWRWF (2c-4), FKKFWKWFRRF(B-33), IKQLLHFFQRF (B-38), RWRRLLKKLHHLLH (α-11), LQLLKQLLKLLKQF (a-7);attached at the C- or N-terminus, and a linker selected from L1, SGG(L2), L3 and LC. MIC assays were conducted against a panel of bacteria,including S. mutans (S. mu), S. gordonii (S. gor), S. sanguinis (S.san), and S. mitis (S. mit), to gauge differences in activity betweenattaching the targeting peptide to the C- or N-terminus of the AMPregion or differences in activity between the linker used.

FIG. 19 shows the killing kinetics of selected peptides against oralstreptococci. All tested bacteria, (A) S. mitis (Smit), (B) S. gordonii(Sgor), (C) S. sanguinis (Ssan), and (D) S. mutans (Smut), were treatedwith peptide solutions at 25 μg/mL for 30 s to 2 h and survivors plated.Data represent an average of three independent experiments.

FIG. 20 illustrates the inhibitory activity of STAMPs against S. mutansbiofilms. S mutans monoculture biofilms were grown and exposed to a 25μg/mL of STAMP, unmodified parental AMP or oral antiseptic (for 1 min),washed, and replenished with fresh medium. Biofilm recovery wasmonitored after 4 h by A600. Data represent an average of threeexperiments.

DETAILED DESCRIPTION

In various embodiments this invention is based on the development of amethod for the rapid identification and synthesis of small peptides fromsequenced genomes for the purposes of quickly screening these moleculesfor diverse biological activities. Previously, small secreted peptideshad to be identified by direct collection (purification of spentmicrobial growth medium or biological tissue/fluid), or byidentification through differential microarray analysis or as aby-product of genetic operon characterization. Both are multi-stepprocesses that terminate at peptide identification; several separateenterprises are required to synthesize or sequence any peptides forcharacterization. Additionally, because of their small size and lack ofprotein domain homologies, small peptides are frequently overlooked whendescribing biological systems, though many organisms contain numerouspeptide-sized open reading frames (ORFs).

Genomic sequence analysis tools already in the public domain andhigh-throughput solid phase peptide chemistry were used to rapidlyidentify and synthesize biologically active peptides. Using sequencesand sometimes annotated genomes, genomes were scanned, by hand or with asearch algorithm, for ORFs predicted to encode peptides of less than forexample, 50-60 amino acids (ignoring small tRNA ORFs or other wellcharacterized genes).

Once noted, these peptides were batch synthesized on a multiplex peptidesynthesizer, yielding 5-10 mg of each peptide that could readily bescreened for biological activity (e.g., binding activity and/orantimicrobial activity.

Accordingly, in certain embodiments, this invention pertains to theidentification of novel peptides (see, e.g., Table 2) that specificallyor preferentially bind particular microorganisms (e.g., bacteria) and/orthat have antimicrobial activity. In certain embodiments these peptidescan be attached to effectors (e.g., drugs, labels, etc.) and used astargeting moieties thereby providing a chimeric moiety thatpreferentially or specifically delivers the effector to a targetmicroorganism, a population of target microorganisms, a microbial filmcomprising the target microorganism(s), a biofilm comprising the targetmicroorganism(s), and the like.

In certain embodiments these peptides can be exploited for theirantimicrobial activity to inhibit the growth or proliferation of amicroorganism and/or to inhibit the formation and/or growth of a biofilmcomprising the microorganism. These antimicrobial peptides can be usedalone, in conjunction with other agents (e.g., antibacterial agents),and/or they can be coupled to a targeting moiety to provide a chimericmoiety that preferentially directs the antimicrobial peptide to a targettissue and/or to a target organism (e.g., a bacterium, a population oftarget microorganisms, a microbial film, a biofilm, and the like).

In certain embodiments the antimicrobial peptides and/or the chimericmoieties described herein can be formulated with a pharmaceuticallyacceptable carrier to form a pharmacological composition.

The amino acid sequences of illustrative peptides of this inventionhaving antimicrobial and/or targeting activity against methicillinresistant Streptococcus mutans, Streptococcus pyogenes, and/or Treponemadenticola is shown in Table 2. As with the other peptides describedherein, it will be recognized that these peptides can comprise all “L”form residues, all “D” form residues, mixtures of “L” and “D” residues,and beta peptide sequences. It will also be appreciated in addition tothe D-form and L-form and beta-peptide sequences this invention alsocontemplates retro and retro-inverso forms of each of these peptides. Inretro forms, the direction of the sequence is reversed. In inverseforms, the chirality of the constituent amino acids is reversed (i.e., Lform amino acids become D form amino acids and D form amino acids becomeL form amino acids). In the retro-inverso form, both the order and thechirality of the amino acids is reversed. Thus, for example, a retroform of the of the peptide Smu11(MKNLIETVEKFLTYSDEKLEELAKKNQALREEISRQKSK, SEQ ID NO:11) has the sequenceKSKQRSIEERLAQNKKALEELKEDSYTLFKEVTEILNKM (SEQ ID NO:10). Where the Smu11peptide comprises all L amino acids, the inverse form will comprise allD amino acids and the retro-inverso (retro-inverse) form will have thesequence of SEQ ID NO:10 and comprise all D form amino acids. Alsocontemplated are peptides having the amino acid sequences or retro aminoacid sequences of the peptides in Table 2 (or the other tables shownherein) and comprising one, two, three, four, five, six, seven, eight,nine, or ten conservative substitutions, but retaining substantially thesame binding and/or antimicrobial activity. Also contemplated arepeptides having the amino acid sequences or retro amino acid sequencesof the peptides in Table 2 (or other tables herein) and comprising one,two, three, four, five, six, seven, eight, nine, or ten deletions, butretaining substantially the same binding and/or antimicrobial activity.

In various embodiments, chimeric moieties comprising one or more of thetargeting peptides found in Table 2 attached to one or more effectors(e.g., antimicrobial peptides as described herein) are contemplated. Invarious embodiments, one or more of the antimicrobial peptides found inTable 2 used as effectors or attached to one or more targeting moieties(e.g., targeting peptides, targeting antibodies, and the like) to formchimeric moieties are contemplated.

TABLE 2 Peptides having antimicrobial and/or targeting activity againstone or more of the following: Streptococcus mutans, Streptococcuspyogenes, and Treponema denticola. SEQ ID Amino Acid Sequence ID NOSmu11 MKNLIETVEKFLTYSDEKLEELAKKNQALREEISRQKSK 11 Smu.18GGRAGRIKKLSQKEAEPFEN 12 Smu41 MFIRSKLRRVDFSGVRRGNKHFLLDKLLITLVK 13 Smu68MSALFYDTLAAIWISIAGVDARWGH 14 Smu150GGKVSGGEAVAAIGICATASAAIGGLAGATLVTPYCVGTWGLI 15 RSH Smu151DKQAADTFLSAVGGAASGFTYCASNGVWHPYILAGCAGVGA 16 VGSVVFPH Smu223cGGKYLFLASKTKEYFKSHFREIMIDV 17 Smu225cMFISFVDCIQNIEKIEKELLKIGITDIQINQDAGWLY 18 Smu277YLTEIEGEGLGLGICLGLVGFAGGFAHGVVQGAGVGTAIEPGY 19GTIIGALVDGVGQDLIYGGAGFAAGYSL Smu283GGFDVKGVAASYLAMGTAALGGLACTTPVGAVLYLGAEVCA 20 GAAVIYYGAN Smu299cGGGLYDGANGYAYRDSQGHWAYKVTKTPAQALTDVVVNSW 21 ASGAASFAAYA Smu390GGRSSYNGFSKICFLKIEHFGSYSYQGR 22 Smu423GGGMIRCALGTAGSAGLGFVGGMGAGTVTLPVVGTVSGAAL 23 GGWSGAAVGAATFC Smu427MEKTYHIDGLKCQGCADNVTKRFSELKKVNDVKVDLDKKEV 24 RITGNPSKWSLKRALKGTNYELGAEISmu444 MSVGMGVIERGSFDFSASAILQKRETKCLKNKPFT 25 Smu451MKMRAGQVVFIYKLILVLLFYVLQKLFDLKKGCF 26 Smu529MLPDSALDERIKGRVSAKNSSLLSALIKKLALIIFIG 27 Smu616GGVATYGAATMGLCAVSGPIGWGLGGAYLLTCAAAGGMIGY 28 GAATLD Smu750cMLSNVLSRSVVSPNVDIPNSMVILSPLLISISNYH 29 Smu812MAILTFFMALLFTYLKEKAQILYWPLFLHLMFYFVTA 30 Smu1018MNTNDLLQAFELMGLGMAGVFIVLGILYIVAELLIKIFPVNN 31 Smu1047cMVHLFSFVKLIYYDIMKYSIEEKVFFESPVGEIIQ 32 Smu1131cGGYATAKLTQTKPTMPKNVKKGTPPKGAPEDTPPNGNSNDSS 33 QSDSDSDSNSSNTNSNSSITNGSmu1231c MDKKRVIERIKSFSLRDEVIHFGELCIYWGK 34 Smu1232cGGKNKLVMSDLRQQVTDMGFVNVKTYINSGNLFFQSDCPRAN 35 ISSRFEQFFADHYPFV Smu1358MNNAYRWFFRLTLADNMHRFTTYGKEWQLSFPK 36 Smu1359MKLAGIEKKINLSKRRKLYLENSSLLINYVKVNMSY 37 Smu1368MTEILNFLIAVHDDRKNWKIKHCLSNSSFDFLCSPDSSR 38 Smu1369MPVQKALHVVSAYATDLGICYDQVVTVMIREVKTQLYQIY 39 Smu1372cGGSYQQVYSWVRKFKKDGINGLLDRRGKGLESKLISVVMVAI 40 VLRPV Smu1504cGGMSSGWLSDDFWLKSAIPLLKKRLAKWNETL 41 Smu1505cMAYSLTFQNPNDNLTDEEVAKYMEKITKALTEKIGAEVR 42 Smu1719cMNTFLWILLVIIALLAGLVGGTFIARKQMEKYLEENPPLNEDVI 43RNMMSQMGQKPSEAKVQQVVRQMNKQQKAAKAKAKKKK Smu1750cMIFNRRKFFQYFGLSKEAMVEHIQPFILDIWQIHLF 44 Smu1752c GGWLNAISLYGRIG 45Smu1768c GGHKQLVIEPLVSQNDQLSLIESLSGILSDSETVDVKDYRSERK 46 EERLKKYESLTSmu1808c MTTTQKTYLHIIRELENQDIDLIMRSLTSLT 47 Smu1813MPMTYCGSPRRTDLAVITDEELGQTLEVINHWPRNV 48 Smu1882cGGATDGEIIANRMLQGKATKGEITMYTWNIIQNGWVNSLVSW 49GIGGYNSSIGYSAQGNRGFSNYPYDVSMDSDNSSSSSNTTGGY VNYNQSFNSGW Smu1889cGGLAGAGTGAAVSAPAAEGGGLGPIAGAAIGWDLGAISGAGL 50 GWANFCQ Smu1895cGGMTWAEIGAIVGATIGSFYIPNPVIVPFRVR 51 Smu1896cGGFGWDSIWRGFKCVAGTAGTIGTGALGGSATGGLTLPIIGHV 52 SGGIIGGISGAGVGIASFCSmu1899 MLSISQRTDRVIVMDKGKIIEEGTHSELIAANGFYHHLFNK 53 Smu1902cGGAFYQRKENVISLDPREWLGFNVTEK 54 Smu1903c GGNIFEYFLE 55 Smu1905cGGGRAPRCAALVGASIYDGLAVVGDPVGVAMAAGTIAAGSFC 56 Smu1906cGGCSWKGADKAGFSGGVGGLIGAGGNPVGGVLGIAGGLDAY 57 GELVGGN Smu1907MEQNILNGSYFVLNGKNAKFLLEIDKLTLPDKLATLPVPHQVR 58 Smu1914cGGGRGWNCAAGIALGAGQGYMATAGGTAFLGPYAIGTGAFG 59 AIAGGIGGALNSCG Smu1915GGSGSLSTFFRLFNRSFTQALGK 60 Smu1948GGVFSVLKHTTWPTRKQSWHDFISILEYSAFFALVIFIFDKLLTL 61 GLAELLKRF Smu1968cGGSVLGKHALFILLKAGFKAYELAGAFEGWKGMHLPTEKC 62 Smu1972cGGLVMNDETIYLFTYENGQISYEEDKRDCSKNV 63 Smu2105MRFLKDELSVSVRLQEKSIEALPFRTKIEIEIESDNQIKTL 64 Smu2106cGGASGEKILEKLIHERKCQLTQNRQIVLKTDLNNLMKDFYK 65 Smu2121cGGIILAKAADLAEIERIISEDPFKINEIANYDIIEFCPTKSSKAFEK 66 VLK

Uses of Targeting Moieties.

When exploited for their targeting activity, the novel targetingpeptides described herein (see, e.g., Table 2) can be used topreferentially or specifically deliver an effector to a microorganism(e.g., a bacterium, a fungus, a protozoan, a yeast, an algae, etc.), toa bacterial film comprising the microorganism, to a biofilm comprisingthe microorganism, and the like. Where the effector comprises an epitopetag and/or a detectable label, binding of the targeting moiety providesan indication of the presence and/or location, and/or quantity of thetarget (e.g., target microorganism). Thus targeting moieties are thusreadily adapted for use in in vivo diagnostics, and/or ex vivo assays.Moreover, because of small size and good stability, the microorganismbinding peptides are well suited for microassay systems (e.g.,microfluidic assays (Lab on a Chip), microarray assays, and the like).

In certain embodiments the microorganism binding peptides (targetingpeptides) can be attached to an effector that has antimicrobial activity(e.g., an antimicrobial peptide, an antibacterial and/or antifungal, avehicle that contains an antibacterial or antifungal, etc. In variousembodiments these chimeric moieties can be used in vivo, or ex vivo topreferentially inhibit or kill the target organism(s).

In certain embodiments the targeting peptides can be used in variouspre-targeting protocols. In pre-targeting protocols, a chimeric moleculeis utilized comprising a primary targeting species (e.g. amicroorganism-binding peptide) that specifically binds the desiredtarget (e.g. a bacterium) and an effector that provides a binding sitethat is available for binding by a subsequently administered secondtargeting species. Once sufficient accretion of the primary targetingspecies (the chimeric molecule) is achieved, a second targeting speciescomprising (i) a diagnostic or therapeutic agent and (ii) a secondtargeting moiety, that recognizes the available binding site of theprimary targeting species, is administered.

An illustrative example of a pre-targeting protocol is the biotin-avidinsystem for administering a cytotoxic radionuclide to a tumor. In atypical procedure, a monoclonal antibody targeted against atumor-associated antigen is conjugated to avidin and administered to apatient who has a tumor recognized by the antibody. Then the therapeuticagent, e.g., a chelated radionuclide covalently bound to biotin, isadministered. The radionuclide, via its attached biotin is taken up bythe antibody-avidin conjugate pretargeted at the tumor. Examples ofpre-targeting biotin/avidin protocols are described, for example, inGoodwin et al., U.S. Pat. No. 4,863,713; Goodwin et al. (1988) J. Nucl.Med. 29: 226; Hnatowich et al. (1987) J. Nucl. Med. 28: 1294; Oehr etal. (1988) J. Nucl. Med. 29: 728; Klibanov et al. (1988) J. Nucl. Med.29: 1951; Sinitsyn et al. (1989) J. Nucl. Med. 30: 66; Kalofonos et al.(1990) J. Nucl. Med. 31: 1791; Schechter et al. (1991) Int. J. Cancer48:167; Paganelli et al. (1991) Cancer Res. 51: 5960; Paganelli et al.(1991) Nucl. Med. Commun. 12: 211; Stickney et al. (1991) Cancer Res.51: 6650; and Yuan et al. (1991) Cancer Res. 51:3119.

It will be recognized that the tumor-specific antibody used for cancertreatments can be replaced with a microorganism binding peptide of thepresent invention and similar pre-targeting strategies can be used todirect labels, antibiotics, and the like to the target organism(s).

Three-step pre-targeting protocols in which a clearing agent isadministered after the first targeting composition has localized at thetarget site also have been described. The clearing agent binds andremoves circulating primary conjugate which is not bound at the targetsite, and prevents circulating primary targeting species(antibody-avidin or conjugate, for example) from interfering with thetargeting of active agent species (biotin-active agent conjugate) at thetarget site by competing for the binding sites on the activeagent-conjugate. When antibody-avidin is used as the primary targetingmoiety, excess circulating conjugate can be cleared by injecting abiotinylated polymer such as biotinylated human serum albumin. This typeof agent forms a high molecular weight species with the circulatingavidin-antibody conjugate which is quickly recognized by thehepatobiliary system and deposited primarily in the liver.

Examples of these protocols are disclosed, e.g., in PCT Application No.WO 93/25240; Paganelli et al. (1991) Nucl. Med. Comm., 12: 211-234; Oehret al. (1988) J. Nucl. Med., 29: 728-729; Kalofonos et al. (1990) J.Nucl. Med., 31: 1791-1796; Goodwin et al. (1988) J. Nucl. Med., 29:226-234; and the like).

These applications of microorganism binding peptides of this inventionare intended to be illustrative and not limiting. Using the teachingprovided herein, other uses will be recognized by one of skill in theart.

Uses of Antimicrobial Peptides.

When exploited for their antimicrobial activity, the novel antimicrobialpeptides described herein (see, e.g., Table 2) can be used to inhibitthe growth and/or proliferation of a microbial species and/or the growthor proliferation of a biofilm comprising the microbial species. Invarious embodiments the peptides can be formulated individually, incombination with each other, in combination with other antimicrobialpeptides, and/or in combination with various antibacterial agents toprovide antimicrobial reagents and/or pharmaceuticals.

In various embodiments, the antimicrobial peptides described herein canbe formulated individually, in combination with each other, incombination with other antimicrobial peptides, and/or in combinationwith various antibiotic (e.g., antibacterial) agents in “homehealthcare” formulations. Such formulations include, but are not limitedto toothpaste, mouthwash, tooth whitening strips or solutions, contactlens storage, wetting, or cleaning solutions, dental floss, toothpicks,toothbrush bristles, oral sprays, oral lozenges, nasal sprays,aerosolizers for oral and/or nasal application, wound dressings (e.g.,bandages), and the like.

In various embodiments the antimicrobial peptides described herein canbe formulated individually, in combination with each other, incombination with other antimicrobial peptides, and/or in combinationwith various antibiotic (e.g., antibacterial) agents in various cleaningand/or sterilization formulations for use in the home, workplace,clinic, or hospital.

In certain embodiments the antimicrobial peptides described herein areattached to one or more targeting moieties to specifically and/or topreferentially deliver the peptide(s) to a target (e.g. a targetmicroorganism, biofilm, bacterial film, particular tissue, etc.).

Other possible uses of the targeting and/or antimicrobial peptidesdisclosed herein include, but are not limited to biofilm dispersal,biofilm retention, biofilm formation, anti-biofilm formation, cellagglutination, induction of motility or change in motility type,chemoattractant or chemorepellent, extracellular signal for sporogenesisor other morphological change, induction or inhibition of virulence geneexpression, utilized as extracellular scaffold, adhesin or binding site,induction or suppression of host immune response, induction orsuppression of bacterial/fungal antimicrobial molecule production,quorum-sensing, induction of swarming behavior, apoptosis or necrosisinducing in eukaryotic cells, affecting control of or inducing theinitiation of cell cycle in eukaryotes, in archaea or prokaryotes,induces autolysis or programmed cell death, inhibition of phage/virusattachment or replication, evasion of innate immunity, induction orinhibition of genetic transformation or transduction competence,induction or inhibition of pilus-mediated conjugation, induction orinhibition of mating behavior in bacteria and fungi, induction orinhibition of nodule formation or metabolic compartmentalization, metal,ion, or nutrient binding, acquisition or inhibition of metal, ion, ornutrient binding and acquisition, and the like.

These applications of the peptides described herein are intended to beillustrative and not limiting. Using the teaching provided herein, otheruses will be recognized by one of skill in the art.

Design and Construction of Chimeric Moieties.

In various embodiments this invention provides chimeric moietiescomprising one or more targeting moieties attached to one or moreeffectors. The targeting moieties are typically selected topreferentially bind to a target microorganism (e.g., bacteria, virus,fungi, yeast, alga, protozoan, etc.), or group of microorganisms (e.g.,gram-negative or gram-positive bacteria, particular genus, particularspecies, etc.) In certain embodiments the targeting moiety comprises oneor more of the targeting peptides shown in Table 2, and/or Table 4and/or Table 6. In certain embodiments the targeting moiety comprisesnon-peptide moieties (e.g., antibodies, receptor, receptor ligand,lectin, and the like).

The effector typically comprises one or more moieties whose activity isto be delivered to the target microorganism(s), and/or to a biofilmcomprising the target microorganism(s), and/or to a cell or tissuecomprising the target microorganism(s), and the like. In certainembodiments the targeting moiety comprises one or more antimicrobialpeptide(s) as described herein (see, e.g., antimicrobial peptides inTable 2, and/or Table 8, and/or Table 9, and/or Table 10), an antibiotic(including, but not limited to a steroid antibiotic) (see, e.g., Table7), a detectable label, a porphyrin, a photosensitizing agent, anepitope tag, a lipid or liposome, a nanoparticle, a dendrimer, and thelike.

In certain embodiments one or more targeting moieties are attached to asingle effector (see, e.g. FIG. 12). In certain embodiments one or moreeffectors are attached to a single targeting moiety. In certainembodiments multiple targeting moieties are attached to multipleeffectors. The targeting moiety(s) can be attached directly to theeffector(s) or through a linker. Where the targeting moiety and theeffector comprise peptides the chimeric moiety can be a fusion protein.

Targeting Moieties.

In various embodiments this invention provides targeting moieties thatpreferentially and/or specifically bind to a microorganism (e.g., abacterium, a fungus, a yeast, etc.). One or more such targeting moietiescan be attached to one or more effectors to provide chimeric moietiesthat are capable of delivering the effector(s) to a target (e.g., abacterium, a fungus, a yeast, a biofilm comprising the bacterium orfungus or yeast, etc.).

In various embodiments, targeting moieties include, but are not limitedto peptides that preferentially bind particular microorganisms (e.g.,bacteria, fungi, yeasts, protozoa, algae, viruses, etc.) or groups ofsuch microorganisms, antibodies that bind particular microorganisms orgroups of microorganisms, receptor ligands that bind particularmicroorganisms or groups of microorganisms, porphyrins (e.g.,metalloporphyrins), lectins that bind particular microorganisms orgroups of microorganisms, and the like. As indicated, it will beappreciated that references to microorganisms or groups of microorganisminclude bacteria or groups of bacteria, viruses or groups of viruses,yeasts or groups of yeasts, protozoa or groups of protozoa, viruses orgroups of viruses, and the like.

Targeting Peptides.

In certain embodiments the chimeric constructs described herein comprisea targeting moiety that is or comprises a targeting peptide. Typicallythe targeting peptides bind particular bacteria, and/or fungi, and/oryeasts, and/or algae, and/or viruses and/or bind particular groups ofbacteria, and/or groups of fungi, and/or groups of yeasts, and/or groupsof algae. The targeting peptides provided can be used to effectivelydeliver one or more effectors to or into a target microorganism.Illustrative targeting peptides include, but are not limited to thetargeting peptides found in Table 2.

Other suitable targeting peptides include the peptides that have beenidentified as binding to particular target organisms as shown in Table 4and/or Table 4.

TABLE 3 Illustrative list of novel targeting peptides. SEQ ID IDTarget(s) Targeting Peptide Sequence NO 1T-1 S. mutans YVNYNQSFNSGW 671T-2 S. mutans NIFEYFLE 68 S. sanguinis S. gordonii S. mitis S. oralisV. atypica Lactobacillus casei Saliva-derived biofilms 1T-3 S. mutansVLGIAGGLDAYGELVGGN 69 S. gordonii 1T-4 S. mutans LDAYGELVGGN 70 S.gordonii S. sanguinis S. oralis V. atypica L. casei 1T-5 S. mutansLGPIAGAAIGWDLGAISGAGL 71 S. sanguinis GWANFCQ 1T-6 S. mutansKFINGVLSQFVLERK 72 1T-7 Myxococcus xanthus SQRIIEPVKSPQPYPGFSVS 73 1T-8M. xanthus FSVAACGEQRAVTFVLLIEDLI 74 1T-9 M. xanthusWAWAESPRCVSTRSNIHALAF 75 RVEVAALT 1T-10 M. xanthus SPAGLPGDGDEA 76 1T-11S. mutans RISE 77 S. epidermidis P. aeruginosa 1T-12 CorynebacteriumFGNIFKGLKDVIETIVKWTAAK 78 xerosis Corynebacterium striatum S.epidermidis S. mutans 1T-13 S. aureus FRSPCINNNSLQPPGVYPAR 79 S.epidermidis P. aeruginosa 1T-14 S. mutans ALAGLAGLISGK 80 S. aureus S.epidermidis C. xerosis 1T-15 S. mutans DVILRVEAQ 81 1T-16 P. aeruginosaIDMR 82 1T-17 S. mutans NNAIVYIS 83 1T-18 S. aureus YSKTLHFAD 84 S.epidermidis C. striatum P. aeruginosa 1T-19 S. aureus PGAFRNPQMPRG 85 S.epidermidis P. aeruginosa 1T-20 S. mutans PALVDLSNKEAVWAVLDDHS 86 P.aeruginosa 1T-21 S. mutans YVEEAVRAALKKEARISTEDTP 87 P. aeruginosaVNLPSFDC 1T-22 S. epidermidis VPLDDGTRRPEVARNRDKDRED 88 P. aeruginosa1T-23 S. mutans PALVDLSNKEAVWAVLDDHS 89 P. aeruginosa 1T-24 P.aeruginosa EEAEEKLAEVSQAVKRLVR 90 1T-25 S. aureus VGLDVSVLVLFFGLQLLSVLLG91 S. epidermidis AMIR C. xerosis C. striatum P. aeruginosa 1T-26 S.mutans LTILPTTFFAIIVPILAVAFIAYSG 92 S. aureus FKIKGIVEHKDQW S.epidermidis Corynebacterium jeikeium C. xerosis C. striatum P.aeruginosa 1T-27 S. mutans ALFVSLEQFLVVVAKSVFALCH 93 S. aureus SGTLS S.epidermidis C. jeikeium C. xerosis C. striatum P. aeruginosa 1T-28 P.aeruginosa VSRDEAMEFIDREWTTLQPAG 94 KSHA 1T-29 S. mutansGSVIKKRRKRMSKKKHRKMLR 95 S. aureus RTRVQRRKLGK S. epidermidis C.jeikeium C. xerosis C. striatum P. aeruginosa 1T-30 S. aureusGKAKPYQVRQVLRAVDKLETR 96 S. epidermidis RKKGGR C. xerosis C. striatum P.aeruginosa 1T-31 S. mutans NATGTDIGEVTLTLGRFS 97 P. aeruginosa 1T-32 S.mutans VSFLAGWLCLGLAAWRLGNA 98 1T-33 S. aureus VRTLTILVIFIFNYLKSISYKLKQ99 S. epidermidis PFENNLAQSMISI C. jeikeium C. xerosis C. striatum P.aeruginosa 1T-34 S. aureus AFWLNILLTLLGYIPGIVHAVYI 100 S. epidermidisIAKR C. jeikeium C. xerosis C. striatum P. aeruginosa 1T-35 P.aeruginosa EICLTLVFPIRGSYSEAAKFPVPI 101 HIVEDGTVELPK 1T-36 S. aureusVYRHLRFIDGKLVEIRLERK 102 S. epidermidis C. jeikeium C. xerosis C.striatum P. aeruginosa 1T-37 S. mutans YIVGALVILAVAGLIYSMLRKA 103 S.aereus S. epidermidis C. jeikeium C. xerosis C. striatum P. aeruginosa1T-38 S. mutans VMFVLTRGRSPRPMIPAY 104 S. aereus S. epidermidis C.jeikeium C. xerosis C. striatum P. aeruginosa 1T-39 S. mutansFGFCVWMYQLLAGPPGPPA 105 P. aeruginosa 1T-40 S. mutans QRVSLWSEVEHEFR 106P. aeruginosa 1T-41 S. mutans KRGSKIVIAIAVVLIVLAGVWVW 107 S. aureus S.epidermidis C. jeikeium C. striatum P. aeruginosa 1T-42 S. aureusTVLDWLSLALATGLFVYLLVA 108 S. epidermidis LLRADRA C. xerosis C. striatumP. aeruginosa 1T-43 C. jeikium DRCLSVLSWSPPKVSPLI 109 P. aeruginosa1T-44 S. mutans DPALADFAAGMRAQVRT 110 S. aureus S. epidermidis C.jeikeium C. striatum P. aeruginosa 1T-45 S. aureusWTKPSFTDLRLGFEVTLYFANR 111 S. epidermidis C. striatum P. aeruginosa1T-46 S. aureus FSFKQRVMFRKEVERLR 112 S. epidermidis C. jeikeium C.xerosis C. striatum P. aeruginosa 1T-47 S. mutans VIKISVPGQVQMLIP 113 S.epidermidis P. aeruginosa 1T-48 S. aureus KLQVHHGRATHTLLLQPPLCA 114 S.epidermidis PGTIR C. jeikeium C. xerosis C. striatum P. aeruginosa 1T-49S. aureus SLVRIHDQQPWVTRGAFIDAAR 115 S. epidermidis TCS C. jeikeium P.aeruginosa 1T-50 P. aeruginosa HSDEPIPNILFKSDSVH 116 1T-51 S. aureusGKPKRMPAEFIDGYGQALLAGA 117 P. aeruginosa 1T-52 S. aureusDEYPAKLPLSDKGATEPRRH 118 C. xerosis P. aeruginosa 1T-53 P. aeruginosaSDILAEMFEKGELQTLVKDAA 119 AKANA 1T-54 S. epidermidis RWVSCNPSWRIQ 120 C.xerosis C. striatum P. aeruginosa 1T-55 C. xerosis NHKTLKEWKAKWGPEAVESW121 P. aeruginosa ATLLG 1T-56 C. xerosis LALIGAGIWMIRKG 122 P.aeruginosa 1T-57 P. aeruginosa RLEYRRLETQVEENPESGRRPM 123 RG 1T-58 P.aeruginosa CDDLHALERAGKLDALLSA 124 1T-59 S. aureus AVGNNLGKDNDSGHRGKKHR125 S. epidermidis KHKHR P. aeruginosa 1T-60 S. aureusYLTSLGLDAAEQAQGLLTILKG 126 S. epidermidis C. jeikeium C. striatum P.aeruginosa 1T-61 P. aeruginosa HATLLPAVREAISRQLLPALVP 127 RG 1T-62 S.epidermidis GCKGCAQRDPCAEPEPYFRLR 128 P. aeruginosa 1T-63 S. aureusEPLILKELVRNLFLFCYARALR 129 S. epidermidis C. jeikeium C. xerosis C.striatum P. aeruginosa 1T-64 S. aureus QTVHHIHMHVLGQRQMHWPPG 130 S.epidermidis C. jeikeium C. xerosis C. striatum P. aeruginosa 1T-65 S.mutans HARAAVGVAELPRGAAVEVEL 131 S. aureus IAAVRP S. epidermidis C.jeikeium C. xerosis C. striatum P. aeruginosa 1T-66 S. mutansDTDCLSRAYAQRIDELDKQYA 132 S. aureus GIDKPL S. epidermidis C. jeikeium C.xerosis C. striatum P. aeruginosa 1T-67 S. aureus GQRQRLTCGRVSGCSEGPSREA133 S. epidermidis AR C. jeikeium C. xerosis C. striatum P. aeruginosa1T-68 S. mutans GGTKEIVYQRG 134 S. aureus C. jeikeium C. xerosis C.striatum P. aeruginosa 1T-69 S. mutans ILSQEADRKKLF 135 P. aeruginosa1T-70 S. aureus NRQAQGERAHGEQQG 136 C. jeikeium P. aeruginosa 1T-71 P.aeruginosa KIDTNQWPPNKEG 137 1T-72 P. aeruginosa EPTDGVACKER 138 1T-73Streptococcus GWWEELLHETILSKFKITKALE 139 pneumoniae LPIQL 1T-74 S.pneumoniae DIDWGRKISCAAGVAYGAIDG 140 CATTV 1T-75 S. pneumoniaeGVARGLQLGIKTRTQWGAATG 141 AA 1T-76 S. pneumoniae EMRLSKFFRDFILWRKK 1421T-77 S. pneumoniae EMRISRIILDFLFLRKK 143 1T-78 S. pneumoniaeFFKTIFVLILGALGVAAGLYIEK 144 NYIDK 1T-79 S. pneumoniaeFGTPWSITNFWKKNFNDRPDF 145 DSDRRRY 1T-80 S. pneumoniae GGNLGPGFGVIIP 1461T-81 S. pneumoniae AIATGLDIVDGKFDGYLWA 147 1T-82 S. pneumoniaeFGVGVGIALFMAGYAIGKDLR 148 KKFGKSC 1T-83 S. pneumoniaeQKPRKNETFIGYIQRYDIDGNG 149 YQSLPCPQN 1T-84 S. pneumoniaeFRKKRYGLSILLWLNAFTNLVN 150 SIHAFYMTLF 1T-85 A. naeslundii, F. nucleatum,VMASLTWRMRAASASLPTHSR 151 P. gingivalis TDA S. epidermidis, S. gordonii,S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-86 S.mitis, S. oralis, S. salivarious HRKNPVLGVGRRHRAHNVA 152 1T-87 S. mitis,S. mutans, S. oralis EAVGQDLVDAHHP 153 1T-88 UnanalyzedGRLVLEITADEVKALGEALAN 154 AKI 1T-89 S. mitis, S. mutansHEDDKRRGMSVEVLGFEVVQH 155 EE 1T-90 S. gordonii, S. mitis, S. mutans,RNVIGQVL 156 S. oralis, S. sanguinis 1T-91 S. mitis, S. mutans, S.oralis, TSVRPGAAGAAVPAGAAGAA 157 S. sanguinis GAGWRWP 1T-92 S. mitis, S.mutans GQDEGQRRAGVGEGQGVDG 158 1T-93 S. epidermidis, S. gordonii,AMRSVNQA 159 S. mitis, S. mutans, S. oralis, S. sanguinis 1T-94 S.mitis, S. mutans, S. oralis DQVAHSGDMLVQARRRDS 160 1T-95 S. gordonii, S.mitis, S. mutans, GHLLRVGGRVGGVGGVAGAC 161 S. oralis, S. sanguinisAQPFGGQ 1T-96 S. gordonii, S. mitis, S. mutans, VAGACAQPFGGQ 162 S.oralis, S. sanguinis 1T-97 A. naeslundii, F. nucleatum,GVAERNLDRITVAVAIIWTITIV 163 P. gingivalis GLGLVAKLG S. epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-98 A. naeslundii, F. nucleatum, VRSAKAVKALTAAGYTGELVN 164 P.gingivalis VSGGMKAWLGQ S. epidermidis, S. gordonii, S. mitis, S. mutans,S. oralis, S. salivarious, S. sanguinis 1T-99 S. gordonii, S. mitis, S.mutans, MKAWLGQ 165 S. oralis, S. sanguinis 1T-100 S. gordonii, S.mitis, S. mutans LDPLEPRIAPPGDRSHQGAPAC 166 HRDPLRGRSARDAER 1T-101 A.naeslundii, P. gingivalis RLRVGRATDLPLTSFAVGVVR 167 S. epidermidis,NLPDAPAH S. gordonii, S. mitis, S. mutans, S. oralis, S. sanguinis1T-102 A. naeslundii, F. nucleatum, WKRLWPARILAGHSRRRMRW 168 P.gingivalis MVVWRYFAAT S. epidermidis, S. gordonii, S. mitis, S. mutans,S. oralis, S. salivarious, S. sanguinis 1T-103 A. naeslundii, F.nucleatum, AQFYEAIITGYALGAGQRIGQL 169 P. gingivalis S. epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. sanguinis 1T-104 S. mitisRAVAAHLQGRHHGHQVRRQR 170 HGQR 1T-105 S. epidermidis, S. gordonii,GEGLPPPVLHLPPPRMSGR 171 S. mitis, S. mutans, S. oralis 1T-106 S.gordonii, S. mitis, S. mutans, DALRRSRSQGRRHR 172 S. oralis, S.salivarious 1T-107 A. naeslundiiS. epidermidis, SPVPRFTAVGGVSRGSP 173 S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-108 S. gordonii, S. mitis, S. mutans, WGPLGPERPLW 174 S. oralis, S.salivarious, S. sanguinis 1T-109 A. naeslundiiS. epidermidis,VTTNVRQGAGS 175 S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-110 A. naeslundii, P. gingivalisLAAKTAVCVGRAFM 176 S. epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. sanguinis 1T-111 A. naeslundii, F. nucleatum,GRLSRREEDPATSIILLRGAYR 177 P. gingivalis MAVF S. epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-112 S. gordonii SDNDGKLILGTSQ 178 1T-113 S. mitisHGAHQRTGQRLHHHRGRTVSG 179 CRQNPVAGVDPDEHR 1T-114 A. naeslundii, P.gingivalis RQAPGPGLVTITAACSAPGSRSR 180 S. epidermidis, S. gordonii, S.mitis, S. mutans, S. oralis, S. sanguinis 1T-115 A. naeslundii, F.nucleatum, LLIERFSNHH 181 P. gingivalis S. epidermidis, S. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-116 A.naeslundii, P. gingivalis MILHRRRDR 182 S. epidermidis, S. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-117 S.mutans GPGVVGPAPFSRLPAHALNL 183 1T-118 A. naeslundii, F. nucleatum,TASPPAPSDQGLRTAFPATLLIA 184 P. gingivalis LAALARISR S. epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-119 S. gordonii, S. mitis, S. mutans, SPATQKAPTRAQPSRAPVQDC 185 S.oralis GDGRPTAAPDDVERLSPR 1T-120 A. naeslundii, F. nucleatum,DVRDRVDLAGADLCAAHATR 186 P. gingivalis S. epidermidis, S. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-121 S.gordonii, S. mitis, S. mutans, FAKETGFGIGGAQEGWWIIADI 187 S. oralis, S.salivarious, YGPNPF S. sanguinis 1T-122 S. mitis GAIPDPVTHRVDWEEDHQTRP188 SR 1T-123 S. gordonii LVRRNAVAGRSDGLAGAEQLD 189 LVRLQGVL 1T-124 S.mitis, S. mutans, S. oralis LFDERNKIA 190 1T-125 S. epidermidis, S.gordonii, DAITGGNPPLSDTDGLRP 191 S. mutans, S. oralis 1T-126 S.gordonii, S. mitis, S. mutans QGLARPVLRRIPL 192 1T-127 A. naeslundii, F.nucleatum, YDPVPKRKNKNSEGKREE 193 P. gingivalis, T. denticolaS.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-128 A. naeslundii, P. gingivalis SGSAIRMLEIATKMLKR 194 S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-129 A. naeslundii, P. gingivalisYDKYIKYLSIQPPFIVYFI 195 S. epidermidis, S. gordonii, S. mitis, S.mutans, S. oralis, S. salivarious, S. sanguinis 1T-130 A. naeslundii, F.nucleatum, QKIIDMSKFLFSLILFIMIVVIYI 196 P. gingivalisGKSIGGYSAIVSSIMLELDTVLY S. epidermidis, NKKIFFIYK S. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-131 A. naeslundii,F. nucleatum, DEVWKMLGI 197 P. gingivalis, T. denticolaS. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-132 A.naeslundii, F. nucleatum, YSKKLFEYFYFIIFILIRYLIFYKI 198 P. gingivalisIQNKNYYINNIAYN S. epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-133 A. naeslundii, P. gingivalisYFIKDDNEALSKDWEVIGNDL 199 S. epidermidis, KGTIDKYGKEFKVR S. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-134 A.naeslundii, F. nucleatum, SRLVREIKKKCRKS 200 P. gingivalis S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-135 A. naeslundii, P. gingivalisFESLLPQATKKIVNNKGSKINKIF 201 S. epidermidis, S. gordonii, S. mitis, S.mutans, S. oralis, S. salivarious, S. sanguinis 1T-136 A. naeslundii, F.nucleatum, ELLTQIRLALLYSVNEW 202 P. gingivalis, S. epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-137 A. naeslundii, F. nucleatum, PLNFYRAVKENRLPLSEKNIND 203 P.gingivalis FTNIKLKVSPKLINLLQESSIFY S. epidermidis, NFSPKKRNTN S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-138 A. naeslundii, F. nucleatum, YPNEYCIFLENLSLEELKEIKAI 204 P.gingivalis NGETLNLEEIINERKNLKD S. epidermidis, S. gordonii, S. mitis, S.mutans, S. oralis, S. salivarious, S. sanguinis 1T-139 A. naeslundiiS.gordonii, AVAGAAVGALLGNDARSTAV 205 S. mitis, S. mutans, S. oralisGAAIGGALGAGAGELTKNK 1T-140 A. naeslundii, F. nucleatum,IKGTIAFVGEDYVEIRVDKGVK 206 P. gingivalis LTFRKSAIANVINNNQQ S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-141 F. nucleatum, P. gingivalisKKFIILLFILVQGLIFSATKTLSD 207 S. epidermidis, IIAL S. gordonii, S. mitis,S. mutans, S. oralis, S. sanguinis 1T-142 A. naeslundii, F. nucleatum,FTQGIKRIVLKRLKED 208 P. gingivalis, T. denticolaS. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-143 A. naeslundii,F. nucleatum, MPKRHYYKLEAKALQFGLPFA 209 P. gingivalis YSPIQLLK S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-144 A. naeslundii, F. nucleatum,IIELHPKSWTQDWRCSFL 210 P. gingivalis, T. denticolaS. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-145 S. mitis, S.mutans, S. oralis VEAGKRNISLENIEKISKGLGISI 211 SELFKYIEEGEDKIG 1T-146 A.naeslundii, F. nucleatum, RNSADNQTKIDKIRIDISLWDE 212 P. gingivalis,HLNIVKQGK T. denticolaS. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-147 A. naeslundii, F. nucleatum,GVENRRFYERDVSKVSMMTSE 213 P. gingivalis, AVAPRGGSK T. denticolaS.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-148 A. naeslundii, F. nucleatum, IVELDDTTILERALSMLGEANA 214 P.gingivalis, T. denticolaS. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-149 A. naeslundii, F. nucleatum,SVRAVKPIDETVARHFPGDFIVN 215 P. gingivalis, T. denticolaS. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-150 A.naeslundii, F. nucleatum, YINRRLKKAFSDADIKEAPAEF 216 P. gingivalis,YEELRRVQYV T. denticolaS. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-151 A. naeslundii, F. nucleatum,SVRAVKPIDEIVAWHFPGDFIVN 217 P. gingivalis, T. denticolaS. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-152 A.naeslundii, F. nucleatum, YVSADESAYNHIVTDDIPLADR 218 P. gingivalisRIEAVQQ S. epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-153 A. naeslundii, F. nucleatum, YIACPGYFY219 P. gingivalis S. epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-154 P. gingivalis YFSFLEIVGMARR220 1T-155 A. naeslundii, F. nucleatum, LKLAFGVYPFQAMSQSDTAVS 221 P.gingivalis ERNVLWR S. epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-156 A. naeslundii, F. nucleatum,GRFQISIRGEEKSKVKVQGKGT 222 P. gingivalis, FTDRNTT T. denticolaS.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-157 A. naeslundii, F. nucleatum, RRFRKTTENREKSKNKKAVLG 223 P.gingivalis, LSTTSTASY T. denticolaS. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-158 A. naeslundii, F. nucleatum,WENKPSPLGSIKKLQGLVYRLI 224 P. gingivalis GYRHFWV S. epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-159 P. gingivalis IFSLHHFALICSEMGTFAVSKRA 225 KYKWEVL 1T-160 A.naeslundii, F. nucleatum, AQYKYINKLLN 226 P. gingivalis, T. denticolaS.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-161 A. naeslundii, F. nucleatum, NKVLQVEVMWDGSVVGRPAG 227 P.gingivalis VISIKSSKKG S. epidermidis, S. gordonii, S. mitis, S. mutans,S. oralis, S. salivarious, S. sanguinis 1T-162 A. naeslundii, F.nucleatum, QKAKEESDRKAAVSYNGFHRV 228 P. gingivalis, NVVSIPK T.denticolaS. gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S.sanguinis 1T-163 A. naeslundii, F. nucleatum, MENILIYIPMVLSPFGSGILLFLG229 P. gingivalis KDRRYML S. epidermidis, S. gordonii, S. mitis, S.mutans, S. oralis, S. salivarious, S. sanguinis 1T-164 A. naeslundii, F.nucleatum, KKSHSQGKRKLKDLNSAYKID 230 P. gingivalis NQLHYALR S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-165 A. naeslundii, F. nucleatum,CYDSFDFSIFVTFANRMKLSVGS 231 P. gingivalis S. epidermidis, S. gordonii,S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-166 A.naeslundii, F. nucleatum, AQSAGQIKRKSKVRIHV 232 P. gingivalis S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-167 A. naeslundii, F. nucleatum,SRMSEHSPAGLVFEVGPMDKG 233 P. gingivalis SFIILDSYHPTVKK S. epidermidis,S. gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S.sanguinis 1T-168 A. naeslundii, F. nucleatum, ELHRIMSTEKIGAVTKMNFDTA 234P. gingivalis PIMSILPIDIYPKEVGIGS S. epidermidis, S. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-169 A. naeslundii,F. nucleatum, FARVRRLHQNRILTQPLTNLKY 235 P. gingivalis CLRQPIYSD S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-170 P. gingivalis AYGKVFSMDIMLSENDKLIVLR236 ISHHSAWH 1T-171 A. naeslundii, F. nucleatum, SVRAVKPIDKTVARHFPGDFIVN237 P. gingivalis S. epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-172 A. naeslundii, F. nucleatum,FEGLKNLLGDDII 238 P. gingivalis S. epidermidis, S. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-173 A. naeslundii,F. nucleatum, LFRKEDQEHVLL 239 P. gingivalis S. gordonii, S. mitis, S.mutans, S. oralis, S. salivarious, S. sanguinis 1T-174 A. naeslundii, F.nucleatum, SGGSDTDGSSSGEPGSHSGDL 240 P. gingivalis, T. denticolaS.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-175 A. naeslundii, F. nucleatum, GEPGSHSGDL 241 P. gingivalis, S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-176 A. naeslundii, P. gingivalisPVGDIMSGFLRGANQPRFLLDH 242 S. epidermidis, ISFGS S. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-177 P.gingivalisS. gordonii, GTNVPTQILGYSREERFDYEPA 243 S. mitis, S. mutans,S. oralis, PEQR S. salivarious, S. sanguinis 1T-178 A. naeslundii, F.nucleatum, LLASHPERLSLGVFFVYRVLHL 244 P. gingivalis LLENT S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-179 A. naeslundii, F. nucleatum,TCYPLIQRKTDRAYEA 245 P. gingivalis, T. denticolaS. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-180 A. naeslundii,F. nucleatum, VVFGGGDRLV 246 P. gingivalis, T. denticolaS. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-181 A.naeslundii, F. nucleatum, YGKESDP 247 P. gingivalis, T. denticolaS.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-182 A. naeslundii, F. nucleatum, LTASICRQWNDNSTPYQR 248 P.gingivalis, T. denticolaS. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-183 A. naeslundii, F. nucleatum,PLRSFVAEKAEHAFRVVRIADF 249 P. gingivalisS. DFGHS epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-184 A. naeslundii, F. nucleatum, ALLVLNLLLMQFFFGKNM 250 P.gingivalis, T. denticolaS. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-185 A. naeslundii, F. nucleatum,HYHFLLEFGFHKGDYLE 251 P. gingivalis, T. denticolaS. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-186 S. sanguinisLAKKNQALREEISRQKSK 252 1T-187 S. sanguinis RAGRIKKLSQKEAEPFEN 253 1T-188S. sanguinis HRKDVYKK 254 1T-189 F. nucleatum, S. sanguinisFIRSKLRRVDFSGVRRGNKHFL 255 LDKLLITLVK 1T-190 A. naeslundii, F.nucleatum, IQIIVNAFVEKDKTGAVIEVLYA 256 P. gingivalisS.SNNHEKVKAKYEELVAIS epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-191 F. nucleatum, S. sanguinisSALFYDTLAAIWISIAGVDARW 257 GH 1T-192 S. sanguinis ILVLLALQVELDSKFQY 2581T-193 S. sanguinis LMIFDKHANLKYKYGNRSFGV 259 EAIM 1T-194 F. nucleatum,S. sanguinis LAGATLVTPYCVGWGLIRSH 260 1T-195 UnanalyzedAASGFTYCASNGVWHPY 261 1T-196 F. nucleatum, S. sanguinisKPEKEKLDTNTLMKVVNKALS 262 LFDRLLIKFGA 1T-197 A. naeslundii, F.nucleatum, TEILNFLITVCADRENWKIKHG 263 P. gingivalisS.LSDSVLLIFFARFTGAEYW epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-198 P. gingivalisS.MPVSKKRYMLSSAYATALGIC 264 epidermidis, S. gordonii,YGQVATDEKESEITAIPDLLDY S. mitis, S. mutans, S. oralis, LSVEEYLL S.sanguinis 1T-199 S. sanguinis RAGRIKKLSQKEAEPFEN 265 1T-200 A.naeslundii, F. nucleatumS. MRFKRFDRDYALSGDNVFEVL 266 epidermidis, S.gordonii, TASCDVIERNLSYREMCGLMQ S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-201 S. sanguinis KRKHENVIVAEEMRVIKN 2671T-202 A. naeslundii, F. nucleatum, LCRLEKLCKQFLRQDKVVTYY 268 P.gingivalisS. LMLPYKRAIEAFYQELKERS epidermidis, S. gordonii, S. mitis, S.mutans, S. oralis, S. salivarious, S. sanguinis 1T-203 A. naeslundii, F.nucleatum, YPFCLATVDHLPEGLSVTDYER 269 P. gingivalisS. VQRLVSQFLLNKEERepidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-204 F. nucleatum, S. sanguinisKYLFLASKTKEYFKSHFREIMI 270 DV 1T-205 A. naeslundii, F. nucleatum,FISFVDCIQNIEKIEKELLKIGIT 271 P. gingivalisS. DIQINQDAGWLY epidermidis,S. gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S.sanguinis 1T-206 S. sanguinis AGFAAGYSL 272 1T-207 F. nucleatum, S.sanguinis SPLEKYGTGSMTALTFLLGCCL 273 LVLSKKSR 1T-208 UnanalyzedKRKRWAILTLFLAGLGAVGIVL 274 ATF 1T-209 F. nucleatum, S. sanguinisWSGAAVGAATFC 275 1T-210 S. sanguinis SVGMGVIERGSFDFSASAILQK 276RETKCLKNKPFT 1T-211 S. sanguinis AEPIIKVTEG 277 1T-212 S. sanguinisLLSALIKKLALIIFIG 278 1T-213 S. sanguinis AYLLTCAAAGGMIGYGAATLD 2791T-214 S. sanguinis MIGYGAATLD 280 1T-215 S. sanguinisVCFKDISVFLSPFRGQEVLFCG 281 KAKHSLIYVIGT 1T-216 S. sanguinisFFLNVIAIRIPHF 282 1T-217 F. nucleatum, S. sanguinisMLSNVLSRSVVSPNVDIPNSMV 283 ILSPLLISISNYH 1T-218 F. nucleatum, S.sanguinis KLIFAALGLVFLLIGLRDSRSK 284 1T-219 S. sanguinisRNINVSATFITEKSLV 285 1T-220 A. naeslundii, F. nucleatum,AILTFFMALLFTYLKEKAQILY 286 P. gingivalisS. WPLFLHLMFYFVTA epidermidis,S. gordonii, S. oralis, S. salivarious, S. sanguinis 1T-221 A.naeslundii, F. nucleatum, DIGRIIGKKGRTITAIRSIVYSVP 287 P. gingivalisS.TQGKKVRLVIDEK epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis,S. salivarious, S. sanguinis 1T-222 F. nucleatum, S. sanguinisRIEASLISAIMFSMFNAIVKFLQK 288 1T-223 A. naeslundii, F. nucleatum,NQKMEINSMTSEKEKMLAGHF 289 P. gingivalisS. HNEANFAVIFKYSLFYNFFepidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-224 A. naeslundii, F. nucleatum,VHLFSFVKLIYYDIMKYSIEEK 290 P. gingivalisS. VFFESPVGEIIQ epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-225 A. naeslundii, F. nucleatum, RRSLGNSASFAEWIEYIRYLHYI 291 P.gingivalisS. IRVQFIHFFSKNKKI epidermidis, S. gordonii, S. mitis, S.mutans, S. oralis, S. salivarious, S. sanguinis 1T-226 A. naeslundii, F.nucleatumS. KLQEKQIDRNFERVSGYSTYRA 292 epidermidis, VQAAKAKEKGFISLEN S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-227 S. sanguinis RFEQFFADHYPFV 293 1T-228 A. naeslundii, F.nucleatum, IFKLFEEHLLYLLDAFYYSKIFR 294 P. gingivalisS.RLKQGLYRRKEQPYTQDLFRM epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-229 S. gordonii, S. oralis, S.sanguinis LLINYVKVNMSY 295 1T-230 A. naeslundii, F. nucleatum,EFLEKFKVLKQPRKANNISKNR 296 P. gingivalisS. VAMIFLTIHKSRGFLSSPYepidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-231 S. gordonii, S. mitis, S. mutans,FDFLCSPDSSR 297 S. oralis, S. salivarious 1T-232 S. sanguinisAYSLTFQNPNDNLTDEEVAKY 298 MEKITKALTEKIGAEVR 1T-233 A. naeslundii, P.gingivalisS. TDQELEHLIVTELESKRLDFTYS 299 epidermidis, KDITEFFDEAFPEYDQNYS. gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S.sanguinis 1T-234 A. naeslundii, F. nucleatum, DNFYLILKMEERGKSKKTSQTR 300P. gingivalisS. GFRAFFDIIRKKIKKEDGK epidermidis, S. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-235 S. sanguinisGWLSDDFWLKSAIPLLKKRLA 301 KWNETL 1T-236 A. naeslundii, F. nucleatum,PVQKALHVVSAYATDLGICYD 302 P. gingivalisS. QVVTVMIREVKTQLYQIYepidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S. sanguinis1T-237 S. sanguinis EDPVPNHFTLRRNKKEKPSKS 303 1T-238 A. naeslundii, F.nucleatum, IFNRRKFFQYFGLSKEAMVEHI 304 P. gingivalisS. QPFILDIWQIHLFepidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-239 A. naeslundiiS. gordonii,ADDLLNKRLTDLIMENAETVK 305 S. mitis, S. mutans, S. oralis, TIDLDNSD S.sanguinis 1T-240 A. naeslundii, F. nucleatum, VILGNGISNIAQTLGQLPNIAW 306P. gingivalisS. VWIYMVLIAALLEESNVC epidermidis, S. gordonii, S. mitis,S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-241 S. sanguinisTQKTYLHIIRELENQDIDLIMRS 307 LTSLT 1T-242 F. nucleatum, S. sanguinisKQVQNTTLIICGTVLLGILFKSY 308 LKSQKSV 1T-243 A. naeslundii, P.gingivalisS. SENIARFAAAFENEQVVSYAR 309 epidermidis, WFRRSWRGSGSSSRF S.gordonii, S. mitis, S. mutants, S. oralis, S. salivarious, S. sanguinis1T-244 A. naeslundii, P. gingivalis MTWAEIGAIVGATIGSFYIPNPV 310 S.epidermidis, IVPFRVR S. gordonii, S. mitis, S. mutans, S. oralis, S.sanguinis 1T-245 F. nucleatum, S. sanguinis IIGGISGAGVGIASFC 311 1T-246S. sanguinis ISGAGVGIASFC 312 1T-247 A. naeslundii, F. nucleatum,LSISQRTDRVIVMDKGKIIEEGT 313 P. gingivalis HSELIAANGFYHHLFNK S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-248 S. sanguinis IGGALNSCG 314 1T-249 F.nucleatum, S. sanguinis VFSVLKHTTWPTRKQSWHDFIS 315ILEYSAFFALVIFIFDKLLTLGLA ELLKRF 1T-250 S. mitis, S. mutans, S. oralisLVQGDTILIENHVGTPVKDDGK 316 DCLIIREADVLAVVND 1T-251 S. mitis, S. oralis,S. sanguinis LVMNDETIYLFTYENGQISYEE 317 DKRDCSKNV 1T-252 F. nucleatum,S. sanguinis MKKNLKRFYALVLGFIIGCLFV 318 SILIFIGY 1T-253 A. naeslundii,F. nucleatum, KTKESLTQQEKKFLKDYDRKS 319 P. gingivalisLHHFRDILTYCFILDKLTNK S. epidermidis, S. gordonii, S. mitis, S. mutans,S. oralis, S. salivarious, S. sanguinis 1T-254 S. sanguinisRFLKDELSVSVRLQEKSIEALPF 320 RTKIEIEIESDNQIKTL 1T-255 A. naeslundii, F.nucleatum, LFIVEYKDKASVPGEIDNTYVE 321 P. gingivalis SYTYSDILTEKTLIRYFDS. epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-256 S. sanguinis KGKSLMPLLKQINQWGKLYL 3221T-257 A. naeslundii, F. nucleatum, IILAKAADLAEIERIISEDPFKIN 323 P.gingivalis EIANYDIIEFCPTKSSKAFEKVLK S. epidermidis, S. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-258 A.naeslundii, F. nucleatum, TINIDDKVLDYLKKINSKAITID 324 P. gingivalis,LIGCAS T. denticola, S. mitis, S. mutans, S. oralis 1T-259 F. nucleatum,P. gingivalis, EKLKKILLKLAVCGKAWYTL 325 T. denticola, S. mitis, S.mutans, S. oralis, S. sanguinis 1T-260 A. naeslundii, P. gingivalisNILYFIHDENQWEPQKAEIFRG 326 S. epidermidis, SIKHCAWLSS S. gordonii, S.mitis, S. mutans, S. oralis, S. sanguinis 1T-261 F. nucleatum S. mutans,SFEKNKIENNLKIAQAYIYIKPK 327 S. oralis S. sanguinis PRICQA 1T-262 A.naeslundii, F. nucleatum, LSLPLIVLTKSI 328 P. gingivalis S. epidermidis,S. gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S.sanguinis 1T-263 A. naeslundii, F. nucleatum, FIAVSFTGNPATFKLVIGCKADN329 P. gingivalis S. epidermidis, S. gordonii, S. mitis, S. oralis, S.salivarious, S. sanguinis 1T-264 S. sanguinis LEGKFYMAEDFDKTPECFKDYV 3301T-265 A. naeslundii, F. nucleatum, GMFENLLMINFQIMNDLKIEIV 331 P.gingivalis VKDRICAV S. epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-266 S. sanguinis RAGTWLVVDEIR332 1T-267 A. naeslundii, F. nucleatum, RIKEERKNRSYKFFIWRLFDEK 333 P.gingivalis, TGFI T. denticola, S. mitis, S. mutans, S. oralis S.sanguinis 1T-268 F. nucleatum S. mutans, PITPKKEKCGLGTYAPKNPVFS 334 S.oralis S. sanguinis KSRV 1T-269 F. nucleatum S. mutans, PLYVAAVEKINTAKKH335 S. oralis S. sanguinis 1T-270 F. nucleatum S. mutans, VHEFDIQKILQNR336 S. oralis S. sanguinis 1T-271 A. naeslundii, F. nucleatum,FLIQKFLLIKTFPPYRKKYVVIV 337 P. gingivalis SQTGTA S. epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-272 F. nucleatum S. mutans, QLAPIDKQLKAVKKIAFYESES 338 S. oralis S.sanguinis TAAKAVTVA 1T-273 F. nucleatum, P. gingivalis,YNEPNYKWLESYKIYKQRCED 339 T. denticola, RTGMYYTEET S. mitis, S. mutans,S. oralis 1T-274 F. nucleatum S. mutans, ETTTEINAIKLHRIKQRSPQGTR 340 S.oralis S. sanguinis RVN 1T-275 A. naeslundii, F. nucleatum,QVLKNFSISRRYKINNPFFKILL 341 P. gingivalis, FIQLRTL T. denticola, S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-276 A. naeslundii, F. nucleatum,ILTLLILGSIGFFILKIKLKLGRF 342 P. gingivalis S. epidermidis, S. gordonii,S. mitis, S. mutans, S. oralis, S. sanguinis 1T-277 A. naeslundii, F.nucleatum, IYYMRFVNKPLEKTFFKI 343 P. gingivalis, T. denticola S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-278 A. naeslundii, F. nucleatum, SINSSAGIQPHCLSSSFVLRTKH 344 P.gingivalis, CFY S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-279 A. naeslundii, F. nucleatum, FVLRTKHCFY345 P. gingivalis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-280 A. naeslundii, F. nucleatum,TNNKNKVIIKAIKFKNKDFINL 346 P. gingivalis, DLFIYRR T. denticola S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-281 A. naeslundii, F. nucleatum, KYEKLTKENLFIRNSGNMCVFI 347 P.gingivalis YFLFFG S. epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-282 F. nucleatum, P. gingivalis,ISLVFPAYT 348 S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-283 A. naeslundii, F. nucleatum,LCTKLEDKQRGRIPAELFIISPIK 349 P. gingivalis, ILERNDAL T. denticola, S.epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-284 A. naeslundii, F. nucleatum, FQYYFSLKRV350 P. gingivalis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-285 A. naeslundii, F. nucleatum,FFPYYLADFYKQLKFLNEYQT 351 P. gingivalis, KNKDKVVEFK S. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-286 S.sanguinis LGFFNNKADLVKADTERDNRM 352 SSLKIKDL 1T-287 P. gingivalis, T.denticola KGYPLPFQYRLNNH 353 S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-288 F. nucleatum, S. gordonii,RWVGGEPSADIYLSAKDTKT 354 S. salivarious, S. sanguinis 1T-289 F.nucleatum, P. gingivalis, EPSADIYLSAKDTKT 355 S. gordonii, S. mitis, S.mutans, S. oralis, S. sanguinis 1T-290 A. naeslundii, F. nucleatum,IINQLNLILLRLMEILIL 356 P. gingivalis, S. gordonii, S. mitis, S. mutans,S. oralis, S. salivarious, S. sanguinis 1T-291 A. naeslundii, F.nucleatum, DMKIIKLYIKILSFLFIKYCNKK 357 P. gingivalis, LNSVKLKA T.denticola, S. mitis, S. mutans, S. oralis 1T-292 A. naeslundii, F.nucleatum, IINQLNLILLRLMEILIL 358 P. gingivalis S. epidermidis, S.gordonii, S. mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis1T-293 A. naeslundii, F. nucleatum, HVEDCFLLSNARTTAIHGRANP 359 P.gingivalis ARGEPRTRSE S. epidermidis, S. gordonii, S. mitis, S. mutans,S. oralis, S. salivarious, S. sanguinis 1T-294 T. denticolaYDKIADGVFKIGKRGVL 360 1T-295 S. mitis, S. salivarious, KYKLKKIIL 361 S.sanguinis 1T-296 A. naeslundii, F. nucleatum, EYSQQSFKAKPCSERGVLSP 362P. gingivalis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-297 A. naeslundii, F. nucleatum,RSLRLNNALTKLPKLWYNRIKE 363 T. denticola, AFYAYNDYDK S. mitis, S. mutans,S. oralis 1T-298 A. naeslundii, F. nucleatum, ILNKKPKLPLWKLGKNYFRRF 364P. gingivalis, YVLPTFLA T. denticola S. gordonii, S. mitis, S. mutans,S. oralis, S. salivarious, S. sanguinis 1T-299 A. naeslundii, F.nucleatum SMLTSFLRSKNTRSLKMYKDV 365 S. epidermidis, HF S. gordonii, S.mitis, S. mutans, S. oralis, S. salivarious, S. sanguinis 1T-300 A.naeslundii, F. nucleatum, PLIISKAQIKMSGDILGSCFKLF 366 P. gingivalisYLRPFF S. epidermidis, S. gordonii, S. mitis, S. mutans, S. oralis, S.salivarious, S. sanguinis 1T-301 F. nucleatum, S. gordonii,SKLPRVLDASLKL 367 S. sanguinis 1T-302 A. naeslundii, P. gingivalisIIIILPKIYLVCKTV 368 S. epidermidis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-303 A. naeslundii, F. nucleatum,LDYENMDCKKRIRI 369 P. gingivalis, S. gordonii, S. mitis, S. mutans, S.oralis, S. salivarious, S. sanguinis 1T-304 P. gingivalisSTAGEASRRTASEASRRTAAKL 370 RG TT-305 F. nucleatum ARNALNMRDVPVDAAIIGIIDG371 MDEE TT-306 F. nucleatum KILNEAEGKLLKVIEKNGEIDIE 372 EI TT-307 F.nucleatum NGDKKAKEELDKWDEVIKELN 373 IQF TT-308 F. nucleatumGLVIIPNLIALIILFSQVRQQTKD 374 YFSNPKLSSR TT-309 F. nucleatumEPLPLTKYDKKDTEMKKVFKEI 375 LAGKVGYEKEEE TT-310 F. nucleatumTKLKKNNKLLSAKKENTLHTK 376 DK TT-311 S. mutans, S. sobrinus AIFDAMHNL 377

As described above, in certain embodiments of the present invention, thetargeting moiety can comprise targeting peptide capable of binding,specifically binding, or preferentially binding to a microorganism,e.g., a target microbial organism. In one embodiment, the targetingpeptide be identified via screening peptide libraries. For example, aphage display peptide library can be screened against a target microbialorganism or a desired antigen or epitope thereof. Any peptide identifiedthrough such screening can be used as a targeting peptide for the targetmicrobial organism. Illustrative additional targeting peptides are shownin Table 4.

TABLE 4 Additional illustrative targeting moieties. Targeting Moiety/SEQ ID Organism Structure/sequence NO LPSB-1 RGLRRLGRRGLRRLGR 378 Phob-1KPVLPVLPVLPVL 379 LPSB-2 VLRIIRIAVLRIIRIA 380 LPTG-1 LPETGGSGGSLPETG 381α-1 RAHIRRAHIRR 382 ANION-1 DEDEDDEEDDDEEE 383 PHILIC-1 STMCGSTMCGSTMCG384 SA5.1/S. aureus VRLPLWLPSLNE 385 SA5.3/S. aureus ANYFLPPVLSSS 386SA5.4/S. aureus SHPWNAQRELSV 387 SA5.5/S. aureus SVSVGMRPMPRP 388SA5.6/S. aureus WTPLHPSTNRPP 389 SA5.7/S. aureus SVSVGMKPSPRP 390SA5.8/S. aureus SVSVGMKPSPRP 391 SA5.9/S. aureus SVPVGPYNESQP 392SA5.10/S. aureus WAPPLFRSSLFY 393 SA2.2/S. aureus WAPPXPXSSLFY 394SA2.4/S. aureus HHGWTHHWPPPP 395 SA2.5/S. aureus SYYSLPPIFHIP 396SA2.6/S. aureus HFQENPLSRGGEL 397 SA2.7/S. aureus FSYSPTRAPLNM 398SA2.8/S. aureus SXPXXMKXSXXX 399 SA2.9/S. aureus VSRHQSWHPHDL 400SA2.10/S. aureus DYXYRGLPRXET 401 SA2.11/S. aureus SVSVGMKPSPRP 402 S.aureus/Consensus V/Q/H-P/H-H-E-F/Y-K/H-H/A-L/H-X-X-K/R-P/L 403 DH5.1/E.coli KHLQNRSTGYET 404 DH5.2/E. coli HIHSLSPSKTWP 405 DH5.3/E. coliTITPTDAEMPFL 406 DH5.4/E. coli HLLESGVLERGM 407 DH5.5/E. coliHDRYHIPPLQLH 408 DH5.6/E. coli VNTLQNVRHMAA 409 DH5.7/E. coliSNYMKLRAVSPF 410 DH5.8/E. coli NLQMPYAWRTEF 411 DH5.9/E. coliQKPLTGPHFSLI 412 CSP/S. mutans SGSLSTFFRLFNRSFTQALGK 413 CSPC18/S.mutans LSTFFRLFNRSFTQALGK 414 CSPC16/S. mutans TFFRLFNRSFTQALGK 415CSPM8/S. mutans TFFRLFNR 416 KH/Pseudomonas spp KKHRKHRKHRKH 417 (US2004/0137482) cCF10 LVTLVFV 418 AgrD1 YSTCDFIM 419 AgrD2 GVNACSSLF 420AgrD3 YINCDFLL 421 NisinA ITSISLCTPGCKTGALMGCNMRTATCIICSIIIVSK 422 PlnAKSSAYSLQMGATAIKQVKKLFKKWGW 423 S3L1-5 WWYNWWQDW 424 PenetratinRQIKIWFWNRRMKWKK* 425 Tat EHWSYCDLRPG 426 Pep-1N KETWWETWWTEW 427 Pep27MRKEFHNVLSSGQLLADKRPARDYNRK 428 HABP35 LKQKIKHVVKLKVVVKLRSQLVKRKQN 429HABP42 (all D) STMMSRSHKTRSHHV 430 HABP52 GAHWQFNALTVRGGGS 431 Hi3/17KQRTSIRATEGCLPS 432 α-E. coli peptide QEKIRVRLSA 433 Salivary ReceptorQLKTADLPAGRDETTSFVLV* 434 Adhesion Fragment S1 (Sushi frag.)GFKLKGMARISCLPNGQWSNFPPKCIRECAMVSS 435 (LPS binding) S3 (Sushi frag.)HAEHKVKIGVEQKYGQFPQGTEVTYTCSGNYFLM 436 (LPS binding) MArg.1AMDMYSIEDRYFGGYAPEVG 437 (Mycoplasma infected cell line binding peptideBPI fragment 1 ASQQGTAALQKELKRIKPDYSDSFKIKH 438 (LPS binding) 6,376,462BPI fragment 2 SSQISMVPNVGLKFSISNANIKISGKWKAQKRFLK 439 (LPS binding)6,376,462 BPI fragment 3 VHVHISKSKVGWLIQLFHKKIESALRNK 440 (LPS binding)6,376,462 LBP fragment 1 AAQEGLLALQSELLRITLPDFTGDLRIPH 441 (LPS binding)6,376,462 LBP fragment 2 HSALRPVPGQGLSLSISDSSIRVQGRWKVRKSFFK 442 (LPSbinding) 6,376,462 LBP fragment 3 VEVDMSGDLGWLLNLFHNQIESKFQKV 443 (LPSbinding) 6,376,462 B. anthracis spore ATYPLPIR 444 binding(WO/1999/036081) Bacillus spore binding peptides of 5-12 amino acidscontaining the sequence 445 (WO/1999/036081) Asn-His-Phe-Leu peptides of5-12 amino acids containing the sequence 446 Asn-His-Phe-Leu-ProThr-Ser-Glu-Asn-Val-Arg-Thr (TSQNVRT) 447 A peptide of formulaThr-Tyr-Pro-X-Pro-X-Arg 448 (TYPXPXR) where X is a Ile, Val or Leu. Apeptide having the sequence TSQNVRT. 449 A peptide having the sequenceTYPLPIR 450 LPS binding peptide 1 TFRRLKWK 451 (6,384,188) LPS BP 2(6,384,188) RWKVRKSFFKLQ 452 LPS BP 3 (6,384,188) KWKAQKRFLKMS 453Pseudomonas pilin KCTSDQDEQFIPKGCSK 454 binding peptide (5,494,672)Patents and patent publications disclosing the referenced antibodies areidentified in the table.

In certain embodiments the targeting moieties can comprise otherentities, particularly when utilized with an antimicrobial peptide asdescribed, for example, in Table 2. Illustrative targeting moieties caninclude a polypeptide, a peptide, a small molecule, a ligand, areceptor, an antibody, a protein, or portions thereof that specificallyinteract with a target microbial organism, e.g., the cell surfaceappendages such as flagella and pili, and surface exposed proteins,lipids and polysaccharides of a target microbial organism.

Targeting Antibodies.

In certain embodiments the targeting moieties can comprise one or moreantibodies that bind specifically or preferentially a microorganism orgroup of microorganisms (e.g., bacteria, fungi, yeasts, protozoa,viruses, algae, etc.). The antibodies are selected to bind an epitopecharacteristic or the particular target microorganism(s). In variousembodiments such epitopes or antigens are typically is gram-positive orgram-negative specific, or genus-specific, or species-specific, orstrain specific and located on the surface of a target microbialorganism. The antibody that binds the epitope or antigen can direct ananti-microbial peptide moiety or other effector to the site.Furthermore, in certain embodiments the antibody itself can provideanti-microbial activity in addition to the activity provided by effectormoiety since the antibody may engage an immune system effector (e.g., aT-cell) and thereby elicit an antibody-associated immune response, e.g.,a humoral immune response.

Antibodies that bind particular target microorganisms can be made usingany methods readily available to one skilled in the art. For example, asdescribed in U.S. Pat. No. 6,231,857 (incorporated herein by reference)three monoclonal antibodies, i.e., SWLA1, SWLA2, and SWLA3 have beenmade against S. mutans. Monoclonal antibodies obtained from non-humananimals to be used in a targeting moiety can also be humanized by anymeans available in the art to decrease their immunogenicity and increasetheir ability to elicit anti-microbial immune response of a human.Illustrative microorganisms and/or targets to which antibodies may bedirected are shown, for example, in Tables 5.

Various forms of antibody include, without limitation, whole antibodies,antibody fragments (e.g., (Fab′)₂, Fab′, etc.), single chain antibodies(e.g., scFv), minibodies, Di-miniantibody, Tetra-miniantibody, (scFv)₂,Diabody, scDiabody, Triabody, Tetrabody, Tandem diabody, VHH,nanobodies, affibodies, unibodies, and the like.

Methods of making such antibodies are well known to those of skill inthe art. In various embodiments, such methods typically involveproviding the microorganism, or a component thereof for use as anantigen to raise an immune response in an organism or for use in ascreening protocol (e.g., phage or yeast display).

For example, polyclonal antibodies are typically raised by one or moreinjections (e.g. subcutaneous or intramuscular injections) of the targetmicroorganism(s) or components thereof into a suitable non-human mammal(e.g., mouse, rabbit, rat, etc.).

If desired, the immunizing microorganism or antigen derived therefromcan be administered with or coupled to a carrier protein by conjugationusing techniques that are well-known in the art. Such commonly usedcarriers which are chemically coupled to the peptide include keyholelimpet hemocyanin (KLH), thyroglobulin, bovine serum albumin (BSA), andtetanus toxoid. The coupled peptide is then used to immunize the animal(e.g. a mouse or a rabbit).

The antibodies are then obtained from blood samples taken from themammal. The techniques used to develop polyclonal antibodies are knownin the art (see, e.g., Methods of Enzymology, “Production of AntiseraWith Small Doses of Immunogen: Multiple Intradermal Injections”,Langone, et al. eds. (Acad. Press, 1981)). Polyclonal antibodiesproduced by the animals can be further purified, for example, by bindingto and elution from a matrix to which the peptide to which theantibodies were raised is bound. Those of skill in the art will know ofvarious techniques common in the immunology arts for purification and/orconcentration of polyclonal antibodies, as well as monoclonal antibodiessee, for example, Coligan, et al. (1991) Unit 9, Current Protocols inImmunology, Wiley Interscience).

In certain embodiments the antibodies produced will be monoclonalantibodies (“mAb's”). The general method used for production ofhybridomas secreting mAbs is well known (Kohler and Milstein (1975)Nature, 256:495

Antibody fragments, e.g. single chain antibodies (scFv or others), canalso be produced/selected using phage display and/or yeast displaytechnology. The ability to express antibody fragments on the surface ofviruses that infect bacteria (bacteriophage or phage) or yeasts makes itpossible to isolate a single binding antibody fragment, e.g., from alibrary of greater than 10¹⁰ nonbinding clones. To express antibodyfragments on the surface of phage (phage display) or yeast, an antibodyfragment gene is inserted into the gene encoding a phage surface protein(e.g., pIII) and the antibody fragment-pIII fusion protein is displayedon the phage surface (McCafferty et al. (1990) Nature, 348: 552-554;Hoogenboom et al. (1991) Nucleic Acids Res. 19: 4133-4137).

Since the antibody fragments on the surface of the phage or yeast arefunctional, phage bearing antigen binding antibody fragments can beseparated from non-binding phage by antigen affinity chromatography(McCafferty et al. (1990) Nature, 348: 552-554). Depending on theaffinity of the antibody fragment, enrichment factors of 20fold-1,000,000 fold are obtained for a single round of affinityselection.

Human antibodies can be produced without prior immunization bydisplaying very large and diverse V-gene repertoires on phage (Marks etal. (1991) J. Mol. Biol. 222: 581-597.

In certain embodiments, nanobodies can be used as targeting moieties.Methods of making V_(h)H (nanobodies) are also well known to those ofskill in the art. The Camelidae heavy chain antibodies are found ashomodimers of a single heavy chain, dimerized via their constantregions. The variable domains of these camelidae heavy chain antibodiesare referred to as V_(HH) domains or V_(HH), and can be either used perse as nanobodies and/or as a starting point for obtaining nanobodies.Isolated V_(HH) retain the ability to bind antigen with high specificity(see, e.g., Hamers-Casterman et al. (1993) Nature 363: 446-448). Incertain embodiments such V_(HH) domains, or nucleotide sequencesencoding them, can be derived from antibodies raised in Camelidaespecies, for example in camel, dromedary, llama, alpaca and guanaco.Other species besides Camelidae (e.g. shark, pufferfish) can producefunctional antigen-binding heavy chain antibodies, from which(nucleotide sequences encoding) such naturally occurring V_(HH) can beobtained, e.g. using the methods described in U.S. Patent Publication US2006/0211088.

In various embodiments, for use in therapy, human proteins arepreferred, primarily because they are not as likely to provoke an immuneresponse when administered to a patient. Comparisons of camelid V_(HH)with the V_(H) domains of human antibodies reveals several keydifferences in the framework regions of the camelid V_(HH) domaincorresponding to the V_(H)/V_(L) interface of the human V_(H) domains.Mutation of these human residues to V_(HH) resembling residues has beenperformed to produce “camelized” human V_(H) domains that retain antigenbinding activity, yet have improved expression and solubility.

Libraries of single V_(H) domains have also been derived for examplefrom V_(H) genes amplified from genomic DNA or from mRNA from thespleens of immunized mice and expressed in E. coli (Ward et al. (1989)Nature 341: 544-546) and similar approaches can be performed using theV_(H) domains and/or the V_(L) domains described herein. The isolatedsingle VH domains are called “dAbs” or domain antibodies. A “dAb” is anantibody single variable domain (V_(H) or V_(L)) polypeptide thatspecifically binds antigen. A “dAb” binds antigen independently of otherV domains; however, as the term is used herein, a “dAb” can be presentin a homo- or heteromultimer with other V_(H) or V_(L) domains where theother domains are not required for antigen binding by the dAb, i.e.,where the dAb binds antigen independently of the additional V_(H) orV_(L) domains.

As described in U.S. Patent Publication US 2006/0211088 methods areknown for the cloning and direct screening of immunoglobulin sequences(including but not limited to multivalent polypeptides comprising: twoor more variable domains—or antigen binding domains—and in particularV_(H) domains or V_(HH) domains; fragments of V_(L), V_(H) or V_(HH)domains, such as CDR regions, for example CDR3 regions; antigen-bindingfragments of conventional 4-chain antibodies such as Fab fragments andscFv's, heavy chain antibodies and domain antibodies; and in particularof V_(H) sequences, and more in particular of V_(HH) sequences) that canbe used as part of and/or to construct such nanobodies.

Methods and procedures for the production of VHH/nanobodies can also befound for example in WO 94/04678, WO 96/34103, WO 97/49805, WO 97/49805WO 94/25591, WO 00/43507 WO 01/90190, WO 03/025020, WO 04/062551, WO04/041863, WO 04/041865, WO 04/041862, WO 04/041867, PCT/BE2004/000159,Hamers-Casterman et al. (1993) Nature 363: 446; Riechmann andMuyldermans (1999) J. Immunological Meth., 231: 25-38; Vu et al. (1997)Molecular Immunology, 34(16-17): 1121-1131; Nguyen et al. (2000) EMBOJ., 19(5): 921-930; Arbabi Ghahroudi et al. (19997) FEBS Letters 414:521-526; van der Linden et al. (2000) J. Immunological Meth., 240:185-195; Muyldermans (2001) Rev. Molecular Biotechnology 74: 277-302;Nguyen et al. (2001) Adv. Immunol. 79: 261, and the like, which are allincorporated herein by reference.

In certain embodiments the antibody targeting moiety is a unibody.Unibodies provide an antibody technology that produces a stable, smallerantibody format with an anticipated longer therapeutic window thancertain small antibody formats. In certain embodiments unibodies areproduced from IgG4 antibodies by eliminating the hinge region of theantibody. Unlike the full size IgG4 antibody, the half molecule fragmentis very stable and is termed a uniBody. Halving the IgG4 molecule leftonly one area on the UniBody that can bind to a target. Methods ofproducing unibodies are described in detail in PCT PublicationWO2007/059782, which is incorporated herein by reference in its entirety(see, also, Kolfschoten et al. (2007) Science 317: 1554-1557).

Affibody molecules are class of affinity proteins based on a 58-aminoacid residue protein domain, derived from one of the IgG-binding domainsof staphylococcal protein A. This three helix bundle domain has beenused as a scaffold for the construction of combinatorial phagemidlibraries, from which Affibody variants that target the desiredmolecules can be selected using phage display technology (see, e.g.,Nord et al. (1997) Nat. Biotechnol. 15: 772-777; Ronmark et al. (2002)Eur. J. Biochem., 269: 2647-2655). Details of Affibodies and methods ofproduction are known to those of skill (see, e.g., U.S. Pat. No.5,831,012 which is incorporated herein by reference in its entirety).

It will also be recognized that antibodies can be prepared by any of anumber of commercial services (e.g., Berkeley antibody laboratories,Bethyl Laboratories, Anawa, Eurogenetec, etc.).

Illustrative antibodies that bind various microorganisms are shown inTable 5.

TABLE 5 Illustrative antibodies that bind target microorganisms. SourceAntibody 7,195,763 Polyclonal/monoclonal binds specific Gram(+) cellwall repeats 6,939,543 Antibodies against G(+) LTA 7,169,903 Antibodiesagainst G(+) peptidoglycan 6,231,857 Antibody against S. mutans (Shi)5,484,591 Gram(−) binding antibodies US 2007/0231321 Diabody binding toStreptococcus surface antigen I/II US 2003/0124635 Antibody against S.mutans US 2006/0127372 Antibodies to Actinomyces naeslundii,Lactobacillus casei US 2003/0092086 Antibody to S. sobrinus

In addition, antibodies (targeting moieties) that bind othermicroorganisms can readily be produced using, for example, the methodsdescribed above.

Porphyrins.

In certain embodiments porphyrins, or other photosensitizing agents, canbe used as targeting moieties in the constructs described herein. Inparticular, metalloporphyrins, particularly a number of non-ironmetalloporphyrins mimic heme in their molecular structure and areactively accumulated by bacteria via high affinity heme-uptake systems.The same uptake systems can be used to deliver antibiotic-porphyrin andantibacterial-porphyrin conjugates. Illustrative targeting porphyrinssuitable for this purpose are described in U.S. Pat. No. 6,066,628 andshown herein, for example, in FIGS. 1 and 2.

For example, certain artificial (non-iron) metalloporphyrins (MPs)(Ga-IX, Mn-IX,) are active against Gram-negative and Gram-positivebacteria and acid-fast bacilli (e.g., Y. enterocolitica, N.meningitides, S. marcescens, E. coli, P. mirabills, K. pneumoniae, K.oxytoca, Ps. aeruginosa, C. freundii, E. aerogenes, F. menigosepticum,S. aureus, B. subtilis, S. pyogenes A, E. faecalis, M. smegmatis, M.bovis, M. tuber., S. cerevisiae) as described in Tables 1-5 of U.S. Pat.No. 6,066,628. These MPs can be used as targeting moieties against thesemicroorganisms.

Similarly, some MPs are also growth-inhibitory against yeasts,indicating their usefulness targeting moieties to target Candida species(e.g., Candida albicans, C. krusei, C. pillosus, C. glabrata, etc.) andother mycoses including but not limited to those caused by asTrichophyton, Epidermophyton, Histoplasma, Aspergillus, Cryptococcus,and the like.

Porphyrins, and other photosensitizers, also have antimicrobialactivity. Accordingly, in certain embodiments, the porphyrins, or otherphotosensitizers, can be used as effectors (e.g., attached to targetingpeptides as described herein). In various embodiments the porphyrins orother photosensitizers can provide a dual functionality, e.g., as atargeting moiety and an antimicrobial and can be attached to a targetingpeptide and/or to an antimicrobial peptide as described herein.

Illustrative porphyrins and other photosensitizers are shown in FIGS.1-11 and described in more detail in the discussion of effectors below.

Pheromones.

In certain embodiments, pheromones from microorganisms can be used astargeting moieties. Illustrative pheromones from bacteria and fungi areshown in Table 6. In certain embodiments, chimeric moieties arecontemplated comprising a targeting moiety comprising or consisting ofthe amino acid sequence (or retro or retro-inverso or beta) sequence ofa peptide shown in Table 6 attached to one or more of the peptides shownin Table 2.

TABLE 6 Illustrative bacterial and fungal pheromones utilizable astargeting moieties. Bacterial Pheromones Locus tag Product Sequence SEQID gi|1041118|dbj|BAA11198.1| iPD1 [Enterococcus MKQQKKHIAALLF 455faecalis] ALILTLVS gi|1113947|gb|AAB35253.1| iAM373sex pheromone SIFTLVA456 inhibito [Enterococcus faecalis, Peptide, 7 aa]gi|115412|sp|P13268.1|CAD1_ENTFA Sex pheromone CAD1 LFSLVLAG 457gi|116406|sp|P11932.1|CIA_ENTFA Sex pheromone cAM373 AIFILAS 458(Clumping-inducing agent) (CIA) gi|117240|sp|P13269.1|CPD1_ENTFA Sexpheromone cPD1 FLVMFLSG 459 gi|12056953|gb|AAG48144.1|AF322594_1putative peptide DSIRDVSPTFNKIRR 460 pheromone PrcA WFDGLFK[Lactobacillus paracasei] gi|123988|sp|P24803.1|IAD1_ENTFA Sex pheromoneinhibitor MSKRAMKKIIPLIT 461 determinant precursor LFVVTLVG (iAD1)gi|126362994|emb|CAM35812.1| precursor of pheromone KDEIYWKPS 462peptide ComX [Bacillus amyloliquefaciens FZB42] gi|1587088|prf||2205353Apheromone YSTCDFIM 463 gi|15900442|ref|NP_345046.1| peptide pheromoneBlpC GLWEDLLYNINRY 464 [Streptococcus AHYIT pneumoniae TIGR4]gi|1617436|emb|CAA66791.1| competence pheromone DIRHRINNSIWRDIF 465[Streptococcus gordonii] LKRK gi|1617440|emb|CAA66786.1| competencepheromone DVRSNKIRLWWEN 466 [Streptococcus gordonii] IFFNKKgi|18307870|gb|AAL67728.1|AF456134_2 ComX pheromone PTTREWDG 467precursor [Bacillus mojavensis] gi|18307874|gb|AAL67731.1|AF456135_2ComX pheromone LQIYTNGNWVPS 468 precursor [Bacillus mojavensis]gi|29377808|ref|NP_816936.1| sex pheromone inhibitor MSKRAMKKIIPLIT 469determinant [Enterococcus LFVVTLVG faecalis V583]gi|3342125|gb|AAC27522.1| putative pheromone GAGKNLIYGMGYG 470[Enterococcus faecium] YLRSCNRL gi|41018893|sp|P60242.1|CSP1_STRPNCompetence-stimulating EMRLSKFFRDFILQ 471 peptide type 1 precursor RKK(CSP-1) gi|57489126|gb|AAW51333.1| PcfP [Enterococcus WSEIEINTKQSN 472faecalis] gi|57489152|gb|AAW51349.1| PrgT [Enterococcus HISKERFEAY 473faecalis] gi|58616083|ref|YP_195761.1| UvaF [Enterococcus KYKCSWCKRVYTL474 faecalis] RKDHRTAR gi|58616111|ref|YP_195802.1| PcfP [EnterococcusWSEIEINTKQSN 475 faecalis] gi|58616132|ref|YP_195769.1| PrgQ[Enterococcus MKTTLKKLSRYIA 476 faecalis] VVIAITLIFIgi|58616137|ref|YP_195772.1| PrgT [Enterococcus HISKERFEAY 477 faecalis]gi|6919848|sp|O33689.1|CSP_STROR Competence-stimulating DKRLPYFFKHLFSN478 peptide precursor (CSP) RTK gi|6919849|sp|O33666.1|CSP2_STRMTCompetence-stimulating EMRKPDGALFNLF 479 peptide precursor (CSP) RRRgi|6919850|sp|O33668.1|CSP3_STRMT Competence-stimulating EMRKSNNNFFHFL480 peptide precursor (CSP) RRI gi|6919851|sp|O33672.1|CSP1_STRMTCompetence-stimulating ESRLPKIRFDFIFPR 481 peptide precursor (CSP) KKgi|6919852|sp|O33675.1|CSP4_STRMT Competence-stimulatingEIRQTHNIFFNFFKRR 482 peptide precursor (CSP)gi|6919853|sp|O33690.1|CSP2_STROR Competence-stimulating DWRISETIRNLIFPR483 peptide precursor (CSP) RK gi|999344|gb|AAB34501.1| cOB1bacterialsex VAVLVLGA 484 pheromone [Enterococcus faecalis, Peptide, 8 aa]gi|18307878|gb|AAL67734.1|AF456136_2 ComX pheromone FFEDDKRKSFI 485precursor [Bacillus subtilis] gi|18307882|gb|AAL67737.1|AF456137_2 ComXpheromone FFEDDKRKSFI 486 precursor [Bacillus subtilis]gi|28272731|emb|CAD65660.1| accessory gene regulator MKQKMYEAIAHLF 487protein D, peptide KYVGAKQLVMCC pheromone precursor VGIWFETKIPDELRK[Lactobacillus plantarum WCFS1] gi|28379890|ref|NP_786782.1| accesorygene regulator MKQKMYEAIAHLF 488 protein D, peptide KYVGAKQLVMCCpheromone precursor VGIWFETKIPDELRK [Lactobacillus plantarum WCFS1]gi|57489105|gb|AAW51312.1| PrgF [Enterococcus VVAYVITQVGAIRF 489faecalis] gi|58616090|ref|YP_195779.1| PrgF [Enterococcus VVAYVITQVGAIRF490 faecalis] gi|58616138|ref|YP_195762.1| PrgN [EnterococcusLLKLQDDYLLHLE 491 faecalis] RHRRTKKIIDEN gi|57489117|gb|AAW51324.1| PcfF[Enterococcus EDIKDLTDKVQSLN 492 faecalis] ALVQSELNKLIKRK DQSgi|57489119|gb|AAW51326.1| PcfH [Enterococcus WFLDFSDWLSKVP 493faecalis] SKLWAE gi|58616102|ref|YP_195792.1| PcfF [EnterococcusEDIKDLTDKVQSLN 494 faecalis] ALVQSELNKLIKRK DQSgi|58616104|ref|YP_195794.1| PcfH [Enterococcus WFLDFSDWLSKVP 495faecalis] SKLWAE Fungi 496 gi|1127585|gb|AAA99765.1| mfa1 gene productMLSIFAQTTQTSAS 497 EPQQSPTAPQGRDN GSPIGYSSCVVA gi|1127592|gb|AAA99771.1|mfa2 gene product MLSIFETVAAAAPV 498 TVAETQQASNNEN RGQPGYYCLIAgi|11907715|gb|AAG41298.1| pheromone precursor PSLPSSPPSLLPPLPL 499MFalpha1D LKLLATRRPTLVG [Cryptococcus neoformans MTLCV var. neoformans]gi|13810235|emb|CAC37424.1| M-factor precursor Mfm1 MDSMANSVSSSSV 500[Schizosaccharomyces VNAGNKPAETLNK pombe] TVKNYTPKVPYMC VIAgi|14269436|gb|AAK58071.1|AF378295_1 peptide mating pheromoneMDTFTYVDLAAVA 501 precursor Bbp2-3 AAAVADEVPRDFE [SchizophyllumDQITDYQSYCIIC commune] gi|14269440|gb|AAK58073.1|AF378297_1 peptidemating pheromone SNVHGWCVVA 502 precursor Bbp2-1 [Schizophyllum commune]gi|1813600|gb|AAB41859.1| pheromone precursor NTTAHGWCVVA 503 Bbp1(1)[Schizophyllum commune] gi|24940428|emb|CAD56313.1| a-pheromoneMQPSTVTAAPKDK 504 [Saccharomyces TSAEKKDNYIIKGV paradoxus] FWDPACVIAgi|27549492|gb|AAO17258.1| pheromone phb3.1 GPTWWCVNA 505 [Coprinopsiscinerea] gi|27549494|gb|AAO17259.1| pheromone phb3.2 SGPTWFCIIQ 506[Coprinopsis cinerea] gi|27752314|gb|AAO19469.1| pheromone protein aFTAIFSTLSSSVASK 507 precursor [Cryptococcus TDAPRNEEAYSSG neoformansvar. grubii] NSP gi|2865510|gb|AAC02682.1| MAT-1 pheromoneMFSIFAQPAQTSVS 508 [Ustilago hordei] ETQESPANHGANP GKSGSGLGYSTCV VAgi|3023372|sp|P78742.1|BB11_SCHCO RecName: Full = Mating- NTTAHGWCVVA509 type pheromone BBP1(1); Flags: Precursorgi|3025079|sp|P56508.1|SNA2_YEAST RecName: Full = Protein SDDNYGSLA 510SNA2 gi|37626077|gb|AAQ96360.1| pheromone precursor Phb3 NGLTFWCVIA 511B5 [Coprinopsis cinerea] gi|37626081|gb|AAQ96362.1| pheromone precursorPSWFCVIA 512 Phb3.2 B45 [Coprinopsis cinerea] gi|37626083|gb|AAQ96363.1|pheromone precursor ASWFCTIA 513 Phb3.1 B47 [Coprinopsis cinerea]gi|37961432|gb|AAP57503.1| Ste3-like pheromone PHHKIANASDKRR 514receptor [Thanatephorus RMYFEIFMCAVL cucumeris]gi|400250|sp|P31962.1|MFA1_USTMA RecName: Full = A1- MLSIFAQTTQTSAS 515specific pheromone; EPQQSPTAPQGRDN AltName: Full = Mating GSPIGYSSCVVAfactor A1 gi|400251|sp|P31963.1|MFA2_USTMA RecName: Full = A2-MLSIFETVAAAAPV 516 specific pheromone; TVAETQQASNNEN AltName: Full= Mating RGQPGYYCLIA factor A2 gi|41209131|gb|AAR99617.1| lipopeptidemating SLTYAWCVVA 517 pheromone precursor Bap2(3) [Schizophyllumcommune] gi|41209146|gb|AAR99650.1| lipopeptide mating TSMAHAWCVVA 518pheromone precursor Bap3(2) [Schizophyllum commune]gi|41209149|gb|AAR99653.1| lipopeptide mating GYCVVA 519 pheromoneprecursor Bbp2(8) [Schizophyllum commune] gi|46098187|gb|EAK83420.1|MFA1_USTMA A1- MLSIFAQTTQTSAS 520 SPECIFIC PHEROMONE EPQQSPTAPQGRDN(MATING FACTOR A1) GSPIGYSSCVVA [Ustilago maydis 521]gi|546861|gb|AAB30833.1| M-factor mating MDSMANTVSSSVV 521 pheromoneNTGNKPSETLNKT [Schizosaccharomyces VKNYTPKVPYMCV pombe] IAgi|5917793|gb|AAD56043.1|AF184069_1 pheromone Mfa2 MFSLFETVAAAVK 522[Ustilago hordei] VVSAAEPEHAPTNE GKGEPAPYCIIAgi|6014618|gb|AAF01424.1|AF186389_1 Phb3.2.42 [Coprinus LTWFCVIA 523cinereus] gi|68266363|gb|AAY88882.1| putative pheromone LREKRRRRWFEAF524 receptor STE3.4 MGFGL [Coprinellus disseminatus]gi|71012805|ref|XP_758529.1| A1-specific pheromone MLSIFAQTTQTSAS 525[Ustilago maydis 521] EPQQSPTAPQGRDN GSPIGYSSCVVAgi|72414834|emb|CAI59748.1| mating factor a1.3 MDALTLFAPVSLG 526[Sporisorium reilianum] AVATEQAPVDEER PNRQTFPWIGCVVAgi|72414854|emb|CAI59758.1| mating factor a2.1 MFIFESVVASVQAV 527[Sporisorium reilianum] SVAEQDQTPVSEG RGKPAVYCTIAgi|1127587|gb|AAA99767.1| rba1 gene product PWMSLLFSFLALLA 528LILPKLSKDDPLGL TRQPR gi|151941959|gb|EDN60315.1| pheromone-regulatedASISLIMEGSANIEA 529 membrane protein VGKLVWLAAALPL [Saccharomycescerevisiae AFI YJM789] gi|3025095|sp|Q07549.1|SNA4_YEAST Protein SNA4ARNVYPSVETPLLQ 530 GAAPHDNKQSLVE SPPPYVPgi|73921293|sp|Q08245.3|ZEO1_YEAST RecName: Full = ProteinFLKKLNRKIASIFN 531 ZEO1; AltName: Full = Zeocin resistance protein 1gi|74644573|sp|Q9P305.3|IGO2_YEAST RecName: Full = ProteinDSISRQGSISSGPPP 532 IGO2 RSPNK

Effectors.

Any of a wide number of effectors can be coupled to targeting moietiesas described herein to preferentially deliver the effector to a targetorganism and/or tissue. Illustrative effectors include, but are notlimited to detectable labels, small molecule antibiotics, antimicrobialpeptides, porphyrins or other photosensitizers, epitope tags/antibodiesfor use in a pretargeting protocol, microparticles and/or microcapsules,nanoparticles and/or nanocapsules, “carrier” vehicles including, but notlimited to lipids, liposomes, dendrimers, cholic acid-based peptidemimics or other peptide mimics, steroid antibiotics, and the like.

Detectable Labels.

In certain embodiments chimeric moieties are provided comprising atargeting moiety (e.g. as described in Table 2) attached directly orthrough a linker to a detectable label. Such chimeric moieties areeffective for detecting the presence and/or quantity, and/or location ofthe microorganism(s) to which the targeting moiety is directed.Similarly these chimeric moieties are useful to identify cells and/ortissues and/or food stuffs and/or other compositions that are infectedwith the targeted microorganism(s).

Detectable labels suitable for use in such chimeric moieties include anycomposition detectable by spectroscopic, photochemical, biochemical,immunochemical, electrical, optical, or chemical means. Illustrativeuseful labels include, but are not limited to, biotin for staining withlabeled streptavidin conjugates, avidin or streptavidin for labelingwith biotin conjugates fluorescent dyes (e.g., fluorescein, texas red,rhodamine, green fluorescent protein, and the like, see, e.g., MolecularProbes, Eugene, Oreg., USA), radiolabels (e.g., ³H, ¹²⁵I, ³⁵S, ¹⁴C, ³²P,⁹⁹Tc, ²⁰³Pb, ⁶⁷Ga, ⁶⁸Ga, ⁷²As, ¹¹¹In, ^(113m)In, ⁹⁷Ru, ⁶²Cu, 641Cu,⁵²Fe, ^(52m)Mn, ⁵¹Cr, ¹⁸⁶Re, ¹⁸⁸Re, ⁷⁷As, ⁹⁰Y, ⁶⁷Cu, ¹⁶⁹Er, ¹²¹Sn,¹²⁷Te, ¹⁴²Pr, ¹⁴³Pr, ¹⁹⁸Au, ¹⁹⁹Au, ¹⁶¹Tb, ¹⁰⁹Pd, ¹⁶⁵Dy, ¹⁴⁹Pm, ¹⁵¹Pm,¹⁵³Sm, ¹⁵⁷Gd, ¹⁵⁹Gd, ¹⁶⁶Ho, ¹⁷²Tm, ¹⁶⁹Yb, ¹⁷⁷Lu, ¹⁰⁵Rh, ¹¹¹Ag, and thelike), enzymes (e.g., horse radish peroxidase, alkaline phosphatase andothers commonly used in an ELISA), various colorimetric labels, magneticor paramagnetic labels (e.g., magnetic and/or paramagneticnanoparticles), spin labels, radio-opaque labels, and the like. Patentsteaching the use of such labels include, for example, U.S. Pat. Nos.3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and4,366,241.

It will be recognized that fluorescent labels are not to be limited tosingle species organic molecules, but include inorganic molecules,multi-molecular mixtures of organic and/or inorganic molecules,crystals, heteropolymers, and the like. Thus, for example, CdSe—CdScore-shell nanocrystals enclosed in a silica shell can be easilyderivatized for coupling to a biological molecule (Bruchez et al. (1998)Science, 281: 2013-2016). Similarly, highly fluorescent quantum dots(zinc sulfide-capped cadmium selenide) have been covalently coupled tobiomolecules for use in ultrasensitive biological detection (Warren andNie (1998) Science, 281: 2016-2018).

In various embodiments spin labels are provided by reporter moleculeswith an unpaired electron spin which can be detected by electron spinresonance (ESR) spectroscopy. Illustrative spin labels include organicfree radicals, transitional metal complexes, particularly vanadium,copper, iron, and manganese, and the like. Exemplary spin labelsinclude, for example, nitroxide free radicals.

Means of detecting such labels are well known to those of skill in theart. Thus, for example, where the label is a radioactive label, meansfor detection include a scintillation counter or photographic film as inautoradiography. Where the label is a fluorescent label, it may bedetected by exciting the fluorochrome with the appropriate wavelength oflight and detecting the resulting fluorescence, e.g., by microscopy,visual inspection, via photographic film, by the use of electronicdetectors such as charge coupled devices (CCDs) or photomultipliers andthe like. Similarly, enzymatic labels may be detected by providingappropriate substrates for the enzyme and detecting the resultingreaction product. Finally, simple colorimetric labels may be detectedsimply by observing the color associated with the label.

Antibiotics.

In certain embodiments chimeric moieties are provided comprising atargeting moiety (e.g. as described in Table 2) attached directly orthrough a linker to a small molecule antibiotic and/or to a carrier(e.g., a lipid or liposome, a polymer, etc.) comprising a small moleculeantibiotic (e.g., an antibiotics shown in Table 7).

TABLE 7 Illustrative antibiotics for use in the chimeric moietiesdescribed herein. Class Generic Name Brand Name Aminoglycosides AmikacinAmikin Gentamicin Garamycin Kanamycin Kantrex Neomycin NetilmicinNetromycin Streptomycin Tobramycin Nebcin Paromomycin HumatinCarbacephem Loracarbef Lorabid Carbapenems Ertapenem Invanz DoripenemFinibax Imipenem/Cilastatin Primaxin Meropenem Merrem CephalosporinsCefadroxil Duricef (First Cefazolin Ancef generation) Cefalotin orCefalothin Keflin Cefalexin Keflex Cephalosporins Cefaclor Ceclor(Second Cefamandole Mandole generation) Cefoxitin Mefoxin CefprozilCefzil Cefuroxime Ceftin, Zinnat Cephalosporins Cefixime Suprax (ThirdCefdinir Omnicef generation) Cefditoren Spectracef Cefoperazone CefobidCefotaxime Claforan Cefpodoxime Ceftazidime Fortaz Ceftibuten CedaxCeftizoxime Ceftriaxone Rocephin Cephalosporins Cefepime Maxipime(Fourth generation) Cephalosporins Ceftobiprole (Fifth generation)Glycopeptides Teicoplanin Vancomycin Vancocin Macrolides AzithromycinZithromax Clarithromycin Biaxin Dirithromycin Erythromycin Erythocin,Erythroped Roxithromycin Troleandomycin Telithromycin Ketek MonobactamsAztreonam Penicillins Amoxicillin Novamox, Amoxil Ampicillin AzlocillinCarbenicillin Cloxacillin Dicloxacillin Flucloxacillin FloxapenMezlocillin Meticillin Nafcillin Oxacillin Penicillin PiperacillinTicarcillin Polypeptides Bacitracin Colistin Polymyxin B QuinolonesMafenide Prontosil (archaic) Sulfacetamide Sulfamethizole Sulfanilimide(archaic) Sulfasalazine Sulfisoxazole Trimethoprim Trimethoprim- BactrimSulfamethoxazole (Cotrimoxazole) (TMP-SMX) Tetracyclines DemeclocyclineDoxycycline Vibramycin Minocycline Minocin Oxytetracycline TerracinTetracycline Sumycin Cationic steroid squalamine antibiotics CSA-8CSA-11 CSA-13 CSA-15 CSA-25 CSA-46 CSA-54 CSA-90 CSA-97 OthersArsphenamine Salvarsan Chloramphenicol Chloromycetin Clindamycin CleocinLincomycin Ethambutol Fosfomycin Fusidic acid Fucidin FurazolidoneIsoniazid Linezolid Zyvox Metronidazole Flagyl Mupirocin BactrobanNitrofurantoin Macrodantin, Macrobid Platensimycin PyrazinamideQuinupristin/Dalfopristin Syncercid Rifampin or Rifampicin Tinidazole

Porphyrins and Non-Porphyrin Photosensitizers.

In certain embodiments, porphyrins and other photosensitizers can beused as targeting moieties and/or as effectors in the methods andcompositions of this invention. A photosensitizer is a drug or otherchemical that increases photosensitivity of the organism (e.g.,bacterium, yeast, fungus, etc.). As targeting moieties thephotosensitizers (e.g., porphyrins) are preferentially uptaken by thetarget microorganisms and thereby facilitate delivery of the chimericmoiety to the target microorganism.

As effectors, photosensitizers can be useful in photodynamicantimicrobial chemotherapy (PACT). In various embodiments PACT utilizesphotosensitizers and light (e.g., visible, ultraviolet, infrared, etc.)in order to give a phototoxic response in the target organism(s), oftenvia oxidative damage.

Currently, the major use of PACT is in the disinfection of bloodproducts, particularly for viral inactivation, although moreclinically-based protocols are used, e.g. in the treatment of oralinfection or topical infection. The technique has been shown to beeffective in vitro against bacteria (including drug-resistant strains),yeasts, viruses, parasites, and the like.

Attaching a targeting moiety (e.g., a targeting peptide as shown inTable 2) to the photosensitizer, e.g., as described herein, provides ameans of specifically or preferentially targeting the photosensitizer(s)to particular species or strains(s) of microorganism.

A wide range of photosensitizers, both natural and synthetic are knownto those of skill in the art (see, e.g., Wainwright (1998) J.Antimicrob. Chemotherap. 42: 13-28). Photosensitizers are available withdiffering physicochemical make-up and light-absorption properties. Invarious embodiments photosensitizers are usually aromatic molecules thatare efficient in the formation of long-lived triplet excited states. Interms of the energy absorbed by the aromatic-system, this again dependson the molecular structure involved. For example: furocoumarinphotosensitizers (psoralens) absorb relatively high energy ultraviolet(UV) light (c. 300-350 nm), whereas macrocyclic, heteroaromaticmolecules such as the phthalocyanines absorb lower energy, near-infraredlight.

Illustrative photosensitizers include, but are not limited toporphyrinic macrocyles (especially porphyrins, chlorines, etc., see,e.g., FIGS. 1 and 2). In particular, metalloporphyrins, particularly anumber of non-iron metalloporphyrins mimic heme in their molecularstructure and are actively accumulated by bacteria via high affinityheme-uptake systems. The same uptake systems can be used to deliverantibiotic-porphyrin and antibacterial-porphyrin conjugates.Illustrative targeting porphyrins suitable for this purpose aredescribed in U.S. Pat. No. 6,066,628 and shown herein in FIGS. 1 and 2.

For example, certain artificial (non-iron) metalloporphyrins (MPs)(Ga-IX, Mn-IX,) are active against Gram-negative and Gram-positivebacteria and acid-fast bacilli (e.g., Y. enterocolitica, N.meningitides, S. marcescens, E. coli, P. mirabilis, K. pneumoniae, K.oxytoca, Ps. aeruginosa, C. freundii, E. aerogenes, F. menigosepticum,S. aureus, B. subtilis, S. pyogenes A, E. faecalis, M. smegmatis, M.bovis, M. tuber., S. cerevisiae) as described in Tables 1-5 of U.S. Pat.No. 6,066,628. These MPs can be used as targeting moieties against thesemicroorganisms.

Similarly, some MPs are also growth-inhibitory against yeasts,indicating their usefulness targeting moieties to target Candida species(e.g., Candida albicans, C. krusei, C. pillosus, C. glabrata, etc.) andother mycoses including but not limited to those caused by asTrichophyton, Epidermophyton, Histoplasma, Aspergillus, Cryptococcus,and the like.

Other photosensitizers include, but are not limited to cyanines (see,e.g., FIG. 6) and phthalocyanines (see, e.g., FIG. 4), azines (see,e.g., FIG. 5) including especially methylene blue and toluidine blue,hypericin (see, e.g., FIG. 8), acridines (see, e.g., FIG. 9) includingespecially Rose Bengal (see, e.g., FIG. 10), crown ethers (see, e.g.,FIG. 11), and the like.

In certain embodiments the photosensitizers are toxic or growthinhibitors without light activation. For example, some non-ironmetalloporphyrins (MPs) (see, e.g., FIGS. 1 and 2 herein) possess apowerful light-independent antimicrobial activity. In addition, haemin,the most well known natural porphyrin, possesses a significantantibacterial activity that can augmented by the presence ofphysiological concentrations of hydrogen peroxide or a reducing agent.

Typically, when activated by light, the toxicity or growth inhibitioneffect is substantially increased. Typically, they generate radicalspecies that affect anything within proximity. In certain embodiments toget the best selectivity from targeted photosensitizers, anti-oxidantscan be used to quench un-bound photosensitizers, limiting the damageonly to cells where the conjugates have accumulated due to the targetingpeptide. The membrane structures of the target cell act as the protondonors in this case.

In typical photodynamic antimicrobial chemotherapy (PACT) the targetedphotosensitizer is “activated by the application of a light source(e.g., a visible light source, an ultraviolet light source, an infraredlight source, etc.). PACT applications however need not be limited totopical use. Regions of the mouth, throat, nose, sinuses are readilyilluminated. Similarly regions of the gut can readily be illuminatedusing endoscopic techniques. Other internal regions can be illuminedusing laparoscopic methods or during other surgical procedures. Forexample, in certain embodiments involving the insertion or repair orreplacement of an implantable device (e.g., a prosthetic device) itcontemplated that the device can be coated or otherwise contacted with achimeric moiety comprising a targeting moiety attached to aphotosensitizer as described herein. During the surgical procedureand/or just before closing, the device can be illuminated with anappropriate light source to activate the photosensitizer.

The targeted photosensitizers and uses thereof described herein areillustrative and not to be limiting. Using the teachings providedherein, other targeted photosensitizers and uses thereof will beavailable to one of skill in the art.

Antimicrobial Peptides.

In certain embodiments the chimeric moieties described herein includeone or more antimicrobial peptides (e.g., certain peptides shown inTable 2 and/or Table 10) as effectors. Thus, for example, in certainembodiments, where the peptides described in Table 2 are exploited fortheir targeting ability, chimeric moieties are contemplated comprisingone or more of the targeting peptides of Table 2 attached to one or moreof the antimicrobial peptides of Table 10. In certain embodimentschimeric moieties are contemplated comprising one or more of thetargeting peptides of Table 2 attached to one or more of theantimicrobial peptides of Table 2. In certain embodiments chimericmoieties are contemplated comprising other targeting moieties (e.g.,porphyrins, antibodies, etc.) attached to one or more of theantimicrobial peptides of Table 2.

Antimicrobial peptides (also called host defense peptides) are anevolutionarily conserved component of the innate immune response and arefound among all classes of life. Unmodified, these peptides are potent,broad spectrum antibiotics which demonstrate potential as noveltherapeutic agents. Antimicrobial peptides have been demonstrated tokill Gram-negative and Gram-positive bacteria (including strains thatare resistant to conventional antibiotics), mycobacteria (includingMycobacterium tuberculosis), enveloped viruses, and fungi.

Naturally-occurring antimicrobial peptides are typically short peptides,generally between 12 and 50 amino acids. These peptides often includetwo or more positively charged residues provided by arginine, lysine or,in acidic environments, histidine, and frequently a large proportion(generally >50%) of hydrophobic residues (see, e.g., Papagianni et al.(2003) Biotechnol Adv 21: 465; Sitaram and Nagaraj (2002) Curr Pharm Des8: 727; Dürr et al. (2006) Biochim. Biophys. Acta 1758: 1408-1425).

Frequently the secondary structures of these molecules follow 4 themes,including i) α-helical, ii) β-stranded due to the presence of 2 or moredisulfide bonds, iii) β-hairpin or loop due to the presence of a singledisulfide bond and/or cyclization of the peptide chain, and iv)extended. Many of these peptides are unstructured in free solution, andfold into their final configuration upon partitioning into biologicalmembranes. The ability to associate with membranes is a definitivefeature of antimicrobial peptides although membrane permeabilisation isnot necessary. These peptides have a variety of antimicrobial activitiesranging from membrane permeabilization to action on a range ofcytoplasmic targets.

The modes of action by which antimicrobial peptides kill bacteria isvaried and includes, but is not limited to disrupting membranes,interfering with metabolism, and targeting cytoplasmic components. Inmany cases the exact mechanism of killing is not known.

In various embodiments one or more antimicrobial peptides are used alone(e.g., as broad spectrum antimicrobials) and/or are provided aseffectors attached to one or more targeting moieties thereby providing anarrower spectrum (directed) antimicrobial. In certain embodiments oneor more antimicrobial peptides are provided as effectors attached to oneor more targeting moieties and/or one or more effectors therebyproviding a component of a multiple effector composition/strategy.

Suitable antimicrobial peptides for this use include, but are notlimited to the antimicrobial peptides found in Table 2, and/or Table 9,and/or Table 10.

TABLE 8 Novel antimicrobial peptides. ID Structure/sequence SEQ ID NOK-1 GLGRVIGRLIKQIIWRR 533 K-2 VYRKRKSILKIYAKLKGWH 534 K-3AFYQRKENVISLDPREWLGFNVTEK 535 K-4 DKKRVIERIKSFSLRDEVIHFGELCIYWGK 536 K-5RSSYNGFSKICFLKIEHFGSYSYQGR 537 K-6 WLNAISLYGRIG 538 K-7NYRLVNAIFSKIFKKKFIKF 539 K-8 KIL K FLF K KVF 540 K-9 FI RK FLK KW LL 541K-10 KLFKFLRKHLL 542 K-11 KIL K FLF K QVF 543 K-12 KIL K KLF K FVF 544K-13 GIL K KLF T KVF 545 K-14 L R K FL H K LF 546 K-15 L R KNL R WLF 547K-16 FI RK FLQ KLHL 548 K-17 FTRKFLKFLHL 549 K-18 KKFKKFKVLKIL 550 K-19LLKLLKLKKLKF 551

In certain embodiments peptides that induce alterations in phenotype orother biological activities can also be used as antimicrobial effectormoieties. Illustrative alternative peptides are shown in Table 9.

TABLE 9 Novel growth phenotype-inducing or peptides with other activities. SEQ ID ID Organism, effect Structure/sequence NO G-1S. mutans: Ca2+ DSSQSDSDSDSNSSNTNSNSSITNG 552 bindng G-2S. mutans: biofilm LPGTLHIQAEFPVQLEAGSLIQIFD 553 structure G-3S. mutans: biofilm LACTTPVGAVLYLGAEVCAGAAVI 554 structure YYGAN G-4S. mutans: EIPIQLANDLANYYDISLDSIFFW 555 Biofilm structure G-5M. xanthus: RDMTVAGKRPNFLIITTDEE 556 Altered cell  morphology G-6M. xanthus: NTSIVCAVTFAPIKEVPLLWRAGLT 557 Altered cell  LRSRQSmorphology G-7 M. xanthus: QAKVEREVERDLVYTLRRLCDPSG 558 Altered cell SERTK morphology G-8 S. mutans: PRMIDIISFHGCHGDHQVWTDPQAT 559Altered biofilm  ALPR structure

Other illustrative antimicrobial peptides include, but are not limitedto the AMPs of Table 10.

TABLE 10 Other illustrative antimicrobial peptides. AP numbers refer toID in antimicrobial peptide database (http://aps.unmc.edu/AP/main.php).SEQ ID Effector Structure/Sequence No AP00274 1BH4, Circulin AGIPCGESCVWIPCISAALGCSCKNKVC 560 (CirA, plant YRN cyclotides, XXC, ZZHp)AP00036 1BNB, Beta-defensin 1 DFASCHTNGGICLPNRCPGHMIQIGICF 561 (cow)RPRVKCCRSW AP00047 1BNB, Bovine GPLSCGRNGGVCIPIRCPVPMRQIGTC 562neutrophil beta-defensin FGRPVKCCRSW 12 (BNBD-12, cow) AP00428 1C01,MiAMP1 SAFTVWSGPGCNNRAERYSKCGCSAI 563 (Macadamia integrifoliaHQKGGYDFSYTGQTAALYNQAGCSG antimicrobial peptide 1,VAHTRFGSSARACNPFGWKSIFIQC plant) AP00154 1CIX, Tachystatin A2YSRCQLQGFNCVVRSYGLPTIPCCRGL 564 (Horseshoe crabs, TCRSYFPGSTYGRCQRYCrustacea, BBS) AP00145 1CW5, VNYGNGVSCSKTKCSVNWGQAFQER 565Carnobacteriocin B2 YTAGINSFVSGVASGAGSIGRRP (CnbB2, class IIAbacteriocin, bacteria) AP00153 1CZ6, AndroctoninRSVCRQIKICRRRGGCYYKCTNRPY 566 (scorpions) AP00152 1D6X, TritrpticinVRRFPWWWPFLRR 567 (synthetic) AP00201 1D7N, Mastoparan INLKALAALAKKIL568 (insect) AP00140 1D9J, CecropinA- KWKLFKKIGIGKFLHSAKKF 569 Magainin2hybrid (synthetic) AP00178 1DFN, human alpha DCYCRIPACIAGERRYGTCIYQGRLW570 Defensin HNP-3 AFCC (human neutrophil peptide-3, HNP3, humandefensin, ZZHh) AP01153 1DQC, Tachycitin YLAFRCGRYSPCLDDGPNVNLYSCCS 571(horseshoe crabs, FYNCHKCLARLENCPKGLHYNAYLK Crustacea, BBS) VCDWPSKAGCTAP00437 1DUM, Magainin 2 GIGKYLHSAKKFGKAWVGEIMNS 572 analog (synthetic)AP00451 1E4S, Human beta DHYNCVSSGGQCLYSACPIFTKIQGTC 573 defensin 1(HBD-1, YRGKAKCCK human defensin) AP00149 1EWS, Rabbit kidneyMPCSCKKYCDPWEVIDGSCGLFNSKY 574 defensin 1 (RK-1) ICCREK AP00141 1F0E,CecropinA- KWKLFKKIPKFLHSAKKF 575 Magainin2 Hybrid (P18, synthetic)AP00142 1F0G, CecropinA- KLKLFKKIGIGKFLHSAKKF 576 Magainin2 Hybrid(synthetic) AP00143 1F0H, CecropinA- KAKLFKKIGIGKFLHSAKKF 577 Magainin2Hybrid (synthetic) AP00524 1FD4, Human beta GIGDPVTCLKSGAICHPVFCPRRYKQI578 defensin 2 (HBD-2, GTCGLPGTKCCKKP human defensin, ZZHh) AP004381FJN, Mussel Defensin GFGCPNNYQCHRHCKSIPGRCGGYCG 579 MGD-1 GWHRLPCTCYRCGAP00155 1FRY, SMAP-29 RGLRRLGRKIAHGVKKYGPTVLRIIRI 580 (SMAP29, sheep AGcathelicidin) AP00150 1G89, Indolicidin (cow ILPWKWPWWPWRR 581cathelicidin, BBN, ZZHa) AP00156 1GR4, Microcin J25,VGIGTPISFYGGGAGHVPEYF 582 linear (MccJ25, bacteriocin, bacteria) AP001511HR1, Indolicidin P to ILAWKWAWWAWRR 583 A mutant (synthetic) AP001961HU5, Ovispirin-1 KNLRRIIRKIIHIIKKYG 584 (synthetic) AP00197 1HU6,Novispirin G10 KNLRRIIRKGIHIIKKYG 585 (synthetic) AP00198 1HU7,Novispirin T7 KNLRRITRKIIHIIKKYG 586 (synthetic) AP00445 1HVZ, MonkeyRTD-1 GFCRCLCRRGVCRCICTR 587 (rhesus theta-defensin-1, minidefensin-1,animal defensin, XXC, BBS, lectin, ZZHa) AP00103 1i2v, Heliomicinvariant DKLIGSCVWGAVNYTSDCNGECLLRG 588 (Hel-LL, synthetic)YKGGHCGSFANVNCWCET AP00216 1ICA, Phormia defensinATCDLLSGTGINHSACAAHCLLRGNR 589 A (insect defensin A) GGYCNGKGVCVCRNAP01224 1Jo3, Gramicidin B VGALAVVVWLFLWLW 590 (bacteria) AP01225 1jo4,Gramicidin C VGALAVVVWLYLWLW 591 (bacteria) AP00191 1KFP, Gomesin (Gm,ECRRLCYKQRCVTYCRGR 592 Spider, XXA) AP00283 1KJ6, Huamn betaGIINTLQKYYCRVRGGRCAVLSCLPKE 593 defensin 3 (HBD-3, EQIGKCSTRGRKCCRRKKhuman defensin, ZZHh) AP00147 1KV4, Moricin (insect,AKIPIKAIKTVGKAVGKGLRAINIASTA 594 silk moth) NDVFNFLKPKKRKA AP00227 1L4V,Sapecin (insect, ATCDLLSGTGINHSACAAHCLLRGNR 595 flesh fly)GGYCNGKAVCVCRN AP01161 1L9L, Human GRDYRTCLTIVQKLKKMVDKPTQRSV 596granulysin (huGran) SNAATRVCTRGRSRWRDVCRNFMRRYQSRVIQGLVAGETAQQICEDLRLCIP STGPL AP00026 1LFC, Lactoferricin BFKCRRWQWRMKKLGAPSITCVRRAF 597 (LfcinB, cow, ZZHa) AP00193 1M4F, humanLEAP-1 DTHFPICIFCCGCCHRSKCGMCCKT 598 (Hepcidin 25) AP00499 1MAG,Gramicidin A VGALAVVVWLWLWLW 599 (gA, bacteria) AP00403 1MM0, TermicinACNFQSCWATCQAQHSIYFRRAFCDR 600 (termite defensin, insect SQCKCVFVRGdefensin) AP00194 1MMC, Ac-AMP2 VGECVRGRCPSGMCCSQFGYCGKGP 601 (plantdefensin, BBS) KYCGR AP01206 1MQZ, Mersacidin CTFTLPGGGGVCTLTSECIC 602(bacteria) AP00429 1NKL, Porcine NK- GYFCESCRKIIQKLEDMVGPQPNEDTV 603Lysin (pig) TQAASQVCDKLKILRGLCKKIMRSFL RRISWDILTGKKPQAICVDIKICKE AP00633log7, Sakacin P/ KYYGNGVHCGKHSCTVDWGTAIGNI 604 Sakacin 674 (SakP,GNNAAANWATGGNAGWNK class IIA bacteriocin, bacteria) AP00195 1PG1,Protegrin 1 RGGRLCYCRRRFCVCVGR 605 (Protegrin-1, PG-1, pig cathelicidin,XXA, ZZHa, BBBm) AP00928 1PXQ, Subtilosin A NKGCATCSIGAACLVDGPIPDFEIAGA606 (XXC, class I TGLFGLWG bacteriocin, Gram- positive bacteria) AP004801Q71, Microcin J25 VGIGTPIFSYGGGAGHVPEYF 607 (cyclic MccJ25, class Imicrocins, bacteriocins, Gram-negative bacteria, XXC; BBP) AP00211 1RKK,Polyphemusin I RRWCFRVCYRGFCYRKCR 608 (crabs, Crustacea) AP00430 1T51,IsCT (Scorpion) ILGKIWEGIKSLF 609 AP00731 1ut3, Spheniscin-2SFGLCRLRRGFCARGRCRFPSIPIGRCS 610 (Sphe-2, penguin RFVQCCRRVW defensin,avian defensin) AP00013 1VM5, Aurein 1.2 GLFDIIKKIAESF 611 (frog)AP00214 1WO1, Tachyplesin I KWCFRVCYRGICYRRCR 612 (crabs, Crustacea,XXA, ZZHa) AP00644 1xc0, Pardaxin 4 GFFALIPKIISSPLFKTLLSAVGSALSSS 613(Pardaxin P-4, Pardaxin GGQE P4, Pa4, flat fish) AP00493 1XKM,Distinctin (two NLVSGLIEARKYLEQLHRKLKNCKV 614 chains for stability andtransport? frog) AP00420 1XV3, Penaeidin-4dHSSGYTRPLRKPSRPIFIRPIGCDVCYGI 615 (penaeidin 4, shrimp,PSSTARLCCFRYGDCCHL Crustacea) AP00035 1YTR, Plantaricin AKSSAYSLQMGATAIKQVKKLFKKWGW 616 (PlnA, bacteriocin, bacteria) AP001661Z64, Pleurocidin (fish) GWGSFFKKAAHVGKHVGKAALTHYL 617 AP00780 1Z6V,Human GRRRRSVQWCAVSQPEATKCFQWQR 618 lactoferricinNMRKVRGPPVSCIKRDSPIQCIQA AP00549 1ZFU, Plectasin (fungi,GFGCNGPWDEDDMQCHNHCKSIKGY 619 fungal defensin) KGGYCAKGGFVCKCY AP001771ZMH, human alpha CYCRIPACIAGERRYGTCIYQGRLWAF 620 Defensin HNP-2 CC(human neutrophil peptide-2, HNP2, human defensin, ZZHh) AP00179 1ZMM,human alpha VCSCRLVFCRRTELRVGNCLIGGVSFT 621 Defensin HNP-4 YCCTRVD(human neutrophil peptide-4, HNP4, human defensin) AP00180 1ZMP, humanalpha QARATCYCRTGRCATRESLSGVCEISG 622 Defensin HD-5 (HD5, RLYRLCCR humandefensin) AP00181 1ZMQ, human alpha STRAFTCHCRRSCYSTEYSYGTCTVM 623Defensin HD-6 (HD6, GINHRFCCL human defensin) AP00399 1ZRW, SpinigerinHVDKKVADKVLLLKQLRIMRLLTRL 624 (insect, termite) AP01157 1ZRX, StomoxynRGFRKHFNKLVKKVKHTISETAHVAK 625 (insect) DTAVIAGSGAAVVAAT AP00637 2A2B,Curvacin A/ ARSYGNGVYCNNKKCWVNRGEATQS 626 sakacin A (CurA, SakA,IIGGMISGWASGLAGM class IIA bacteriocin, bacteria) AP00558 2B68, Cg-DefGFGCPGNQLKCNNHCKSISCRAGYCD 627 (Crassostrea gigas AATLWLRCTCTDCNGKKdefensin, oyster defensin, animal defensin) AP01154 2B9K, LCI (bacteria)AIKLVQSPNGNFAASFVLDGTKWIFKS 628 KYYDSSKGYWVGIYEVWDRK AP01005 2DCV,Tachystatin B1 YVSCLFRGARCRVYSGRSCCFGYYCR 629 (BBS, horseshoe crabs)RDFPGSIFGTCSRRNF AP01006 2DCW, Tachystatin B1 YITCLFRGARCRVYSGRSCCFGYYCR630 (BBS, horseshoe crabs) RDFPGSIFGTCSRRNF AP00275 2ERI, Circulin B(CirB, CGESCVFIPCISTLLGCSCKNKVCYRN 631 plant cyclotides, XXC, GVIP ZZHp)AP00707 2f3a, LLAA (LL-37- RLFDKIRQVIRKF 632 derived aurein 1.2 analog,retro-FK13, synthetic) AP00708 2fbs, FK-13 (FK13, FKRIVQRIKDFLR 633NMR-discovered LL-37 core peptide, XXA, ZZHs, synthetic) AP00088 2G9L,Gaegurin-4 GILDTLKQFAKGVGKDLVKGAAQGV 634 (Gaegurin 4, frog) LSTVSCKLAKTCAP01011 2G9P, Latarcin 2a GLFGKLIKKFGRKAISYAVKKARGKH 635 (Ltc2a, BBM,spider) AP00612 2GDL, Fowlicidin-2 LVQRGRFGRFLRKIRRFRPKVTITIQGS 636(chCATH-2, bird ARFG cathelicidin, chicken cathelicidin, BBL) AP004022GL1, VrD2 (Vigna KTCENLANTYRGPCFTTGSCDDHCKN 637 radiata defensin 2,plant KEHLRSGRCRDDFRCWCTRNC defensin, mung bean) AP00285 2GW9,Cryptdin-4 GLLCYCRKGHCKRGERVRGTCGIRFL 638 (Crp4, animal defensin, YCCPRRalpha, mouse) AP00613 2hfr, Fowlicidin-3 RVKRFWPLVPVAINTVAAGINLYKAI 639(chCATH-3, bird RRK cathelicidin, chicken cathelicidin) AP01007 2JMY,CM15 KWKLFKKIGAVLKVL 640 (Synthetic) AP00728 2jni, Arenicin-2 (marineRWCVYAYVRIRGVLVRYRRCW 641 polychaeta, BBBm) AP00473 2jos, Piscidin 1(fish) FFHHIFRGIVHVGKTIHRLVTG 642 AP01151 2JPJ, Lactococcin G-aGTWDDIGQGIGRVAYWVGKALGNLS 643 (chain a, class IIb DVNQASRINRKKKHbacteriocin, bacteria. For chain b, see info) AP00757 2jpy,Phylloseptin-H2 FLSLIPHAINAVSTLVHHF 644 (PLS-H2, Phylloseptin- 2, PS-2)(XXA, frog) AP00546 2jq0, Phylloseptin-1 FLSLIPHAINAVSAIAKHN 645(Phylloseptin-H1, PLS- H1, PS-1, XXA, frog) AP00758 2jq1, Phylloseptin-3FLSLIPHAINAVSALANHG 646 (Phylloseptin-H3, PLS- H3, PS-3) (XXA, frog)AP00727 2jsb, Arenicin-1 (marine RWCVYAYVRVRGVLVRYRRCW 647 polychaeta,BBBm) AP00592 2k10, Ranatuerin-2CSa GILSSFKGVAKGVAKDLAGKLLETLK 648(frog) CKITGC AP00485 2K38, Cupiennin 1a GFGALFKFLAKKVAKTVAKQAAKQG 649(spider) AKYVVNKQME AP00310 2K6O, Human LL-37LLGDFFRKSKEKIGKEFKRIVQRIKDFL 650 (LL37, human RNLVPRTES cathelicidin;released by proteinase 3 from its precursor in neutrophils; FALL-39;BBB, BBM, BBP, BBW, BBD, BBL, ZZHh) AP00199 2LEU, Leucocin AKYYGNGVHCTKSGCSVNWGEAFSAG 651 (LeuA, class IIa VHRLANGGNGFW bacteriocin,bacteria) AP00144 2MAG, Magainin 2 GIGKFLHSAKKFGKAFVGEIMNS 652 (frog)AP00146 2MLT, Melittin (insect, GIGAVLKVLTTGLPALISWIKRKRQQ 653 ZZHa)AP01010 2PCO, Latarcin 1 (Ltc1, SMWSGMWRRKLKKLRNALKKKLKG 654 BBM,spider) EK AP00176 2PM1, human alpha ACYCRIPACIAGERRYGTCIYQGRLW 655Defensin HNP-1 AFCC (human neutrophil peptide-1, HNP1, human defensin,ZZHh) AP01158 2RLG, RP-1 (synthetic) ALYKKFKKKLLKSLKRL 656 AP00102 8TFV,Thanatin (insect) GSKKPVPIIYCNRRTGKCQRM 657 AP00995 A58718, CarnocinUI49 GSEIQPR 658 (bacteria) AP01002 AAC18827, Mutacin IIIKSWSLCTPGCARTGSFNSYCC 659 (mutacin 1140, bacteria) AP00987 ABI74601,Arasin 1 SRWPSPGRPRPFPGRPKPIFRPRPCNCY 660 (Crustacea) APPCPCDRW AP01000CAA63706, variacin GSGVIPTISHECHMNSFQFVFTCCS 661 (lantibiotic, class Ibacteriocin, bacteria) AP00361 O15946, Lebocin 4DLRFWNPREKLPLPTLPPFNPKPIYID 662 (insect, silk moth) MGNRY AP00343O16825, Andropin VFIDILDKMENAIHKAAQAGIGIAKPIE 663 (insect, fruit fly)KMILPK AP00417 O17513, Ceratotoxin D SIGTAVKKAVPIAKKVGKVAIPIAKAV 664(insect, fly) LSVVGQLVG AP00435 O18494, Styelin C (seaGWFGKAFRSVSNFYKKHKTYIHAGLS 665 squirt, tunicate, XXA) AATLL AP00330O18495, Styelin D (Sea GWLRKAAKSVGKFYYKHKYYIKAA 666 squirt, tunicate,XXA) WQIGKHAL AP00331 O18495, Styelin E (Sea GWLRKAAKSVGKFYYKHKYYIKAA667 squirt, tunicate, XXA) WKIGRHAL AP01001 O54329, Mutacin IINRWWQGVVPTVSYECRMNSWQHVF 668 (lantibiotic, mutacin H- TCC 29B, J-T8,class I bacteriocin, bacteria) AP00342 O81338, AntimicrobialAKCIKNGKGCREDQGPPFCCSGFCYR 669 peptide 1 (plant) QVGWARGYCKNR AP00373O96059, Moricin 2 AKIPIKAIKTVGKAVGKGLRAINIASTA 670 (insect)NDVFNFLKPKKRKH AP00449 P01190, Melanotropin SYSMEHFRWGKPV 671 alpha(Alpha-MSH) AP00187 P01376, VVCACRRALCLPRERRAGFCRIRGRIH 672CORTICOSTATIN III PLCCRR (MCP-1, rabbit neutrophil peptide 1, NP-1)(animal defensin, alpha-defensin, rabbit) AP00188 P01377,VVCACRRALCLPLERRAGFCRIRGRIH 673 CORTICOSTATIN IV PLCCRR (MCP-2, rabbitneutrophil defensin 2, NP-2, animal defensin, rabbit) AP00049 P01505,Bombinin GIGALSAKGALKGLAKGLAEHFAN 674 (toad) AP00139 P01507, Cecropin AKWKLFKKIEKVGQNIRDGIIKAGPAVA 675 (insect, ZZHa) VVGQATQIAK AP00128P01509, Cecropin B KWKIFKKIEKVGRNIRNGIIKAGPAVA 676 (insect, silk moth)VLGEAKAL AP00131 P01511, Cecropin D WNPFKELERAGQRVRDAIISAGPAVA 677(insect, moth) TVAQATALAK AP00136 P01518, Crabrolin FLPLILRKIVTAL 678(insect, XXA) AP00183 P04142, Cecropin B RWKIFKKIEKMGRNIRDGIVKAGPAIE 679(insect) VLGSAKAI AP00448 P04205, Mastoparan M INLKAIAALAKKLL 680 (MP-M,insect, XXA) AP00234 P06833, SDEKASPDKHHRFSLSRYAKLANRLA 681Seminalplasmin (SPLN, NPKLLETFLSKWIGDRGNRSV calcium transporterinhibitor, caltrin, cow) AP00314 P07466, RabbitVFCTCRGFLCGSGERASGSCTINGVRH 682 neutrophil peptide 5 TLCCRR (NP-5,animal defensin, alpha-defensin) AP00189 P07467, RabbitVSCTCRRFSCGFGERASGSCTVNGVR 683 neutrophil peptide 4 HTLCCRR (NP-4)AP00186 P07468, GRCVCRKQLLCSYRERRIGDCKIRGV 684 CORTICOSTATIN II RFPFCCPR(Rabbit neutrophil peptide 3b (NP-3b, rabbit) AP00185 P07469,ICACRRRFCPNSERFSGYCRVNGARY 685 CORTICOSTATIN I VRCCSRR (rabbit) AP00217P07469, Rabbit GICACRRRFCPNSERFSGYCRVNGAR 686 neutrophil defensin 3aYVRCCSRR (NP-3a, animal defensin, alpha- defensin) AP00067 P07493,Bombolitin II SKITDILAKLGKVLAHV 687 (insect, bee) AP00068 P07494,Bombolitin III IKIMDILAKLGKVLAHV 688 (insect, bee) AP00069 P07495,Bombolitin IV INIKDILAKLVKVLGHV 689 (insect, bee) AP00070 P07496,Bombolitin V INVLGILGLLGKALSHL 690 (insect, bee) AP00236 P07504,Pyrularia KSCCRNTWARNCYNVCRLPGTISREIC 691 thionin (Pp-TH, plant)AKKCDCKIISGTTCPSDYPK AP00230 P08375, Sarcotoxin IAGWLKKIGKKIERVGQHTRDATIQGLGI 692 (insect, flesh AQQAANVAATAR AP00231P08376, Sarcotoxin IB GWLKKIGKKIERVGQHTRDATIQVIG 693 (insect, fleshVAQQAANVAATAR AP00232 P08377, Sarcotoxin IC GWLRKIGKKIERVGQHTRDATIQVLGI694 (insect, flesh AQQAANVAATAR AP00066 P10521, Bombolitin IIKITTMLAKLGKVLAHV 695 (insect, bee) AP00206 P10946, LantibioticWKSESLCTPGCVTGALQTCFLQTLTC 696 subtilin (class I NCKISK bacteriocin,bacteria) AP00312 P11477, Cryptdin-2 LRDLVCYCRARGCKGRERMNGTCRK 697(Crp2, animal defensin, GHLLYMLCCR alpha, mouse) AP00205 P13068, Nisin AITSISLCTPGCKTGALMGCNMKTATC 698 (lantibiotic, class I HCSIHVSKbacteriocin, bacteria) AP00215 P14214, Tachyplesin II RWCFRVCYRGICYRKCR699 (crabs, Crustacea) AP00212 P14216, Polyphemusin RRWCFRVCYKGFCYRKCR700 II (crabs, Crustacea, XXA, ZZHa. Derivatives: T22) AP00134 P14661,Cecropin P1 SWLSKTAKKLENSAKKRISEGIAIAIQ 701 (pig) GGPR AP00011 P14662,Bactericidin B2 WNPFKELERAGQRVRDAVISAAPAVA 702 (insect) TVGQAAAIARGAP00032 P14663, Bactericidin B- WNPFKELERAGQRVRDAIISAGPAVA 703 3(insect) TVGQAAAIARG AP00033 P14664, Bactericidin B-WNPFKELERAGQRVRDAIISAAPAVA 704 4 (insect) TVGQAAAIARG AP00034 P14665,Bactericidin B- WNPFKELERAGQRVRDAVISAAAVAT 705 5P (insect) VGQAAAIARGAP00125 P14666, Cecropin RWKIFKKIEKVGQNIRDGIVKAGPAV 706 (insect, silkmoth) AVVGQAATI AP00002 P15450, ABAECIN YVPLPNVPQPGRRPFPTFPGQGPFNPKI 707(insect, honeybee) KWPQGY AP00505 P15516, human HistatinDSHAKRHHGYKRKFHEKHHSHRGY 708 5 (ZZHs; derivatives Dh-5) AP00520 P15516,human Histatin 3 DSHAKRHHGYKRKFHEKHHSHRGYR 709 SNYLYDN AP00523 P15516,human Histatin 8 KFHEKHHSHRGY 710 AP00226 P17722, RoyalisinVTCDLLSFKGQVNDSACAANCLSLGK 711 (insect, honeybee)AGGHCEKVGCICRKTSFKDLWDKRF AP00213 P18252, Tachyplesin IIIKWCFRVCYRGICYRKCR 712 (horseshoe crabs, Crustacea) AP00233 P18312,Sarcotoxin ID GWIRDFGKRIERVGQHTRDATIQTIAV 713 (insect, fleshAQQAANVAATLKG AP00207 P19578, Lantibiotic TAGPAIRASVKQCQKTLKATRLFTVS 714PEP5 (class I CKGKNGCK bacteriocin, bacteria) AP00009 P19660, BACTENECINRFRPPIRRPPIRPPFYPPFRPPIRPPIFPPI 715 5 (bac5, cow RPPFRPPLGPFPcathelicidin) AP00010 P19661, BACTENECIN RRIRPRPPRLPRPRPRPLPFPRPGPRPIP716 7 (bac7, cow RPLPFPRPGPRPIPRPLPFPRPGPRPIPRPL cathelicidin) AP00200P21564, Mastoparan B LKLKSIVSWAKKVL 717 (MP-B, insect, XXA) AP00005P21663, Andropin VFIDILDKVENAIHNAAQVGIGFAKPF 718 (insect, fly) EKLINPKAP00008 P22226, Cyclic RLCRIVVIRVCR 719 dodecapeptide (cow cathelicidin)AP01205 P23826, Lactocin S STPVLASVAVSMELLPTASVLYSDVA 720 (XXD3,bacteria) GCFKYSAKHHC AP00239 P24335, XPF (the GWASKIGQTLGKIAKVGLKELIQPK721 xenopsin precursor fragment, African clawed frog) AP00235 P25068,Bovine tracheal NPVSCVRNKGICVPIRCPGSMKQIGTC 722 antimicrobial peptideVGRAVKCCRKK (TAP, cow) AP00418 P25230, CAP18 (rabbitGLRKRLRKFRNKIKEKLKKIGQKIQGF 723 cathelicidin, BBL) VPKLAPRTDY AP00203P25403, Mj-AMP1 QCIGNGGRCNENVGPPYCCSGFCLRQ 724 (MjAMP1, plantPGQGYGYCKNR defensin) AP00202 P25404, Mj-AMP2 CIGNGGRCNENVGPPYCCSGFCLRQP725 (MjAMP2, plant NQGYGVCRNR defensin) AP00138 P28310, Cryptdin-3LRDLVCYCRKRGCKRRERMNGTCRK 726 (Crp3, animal defensin, GHLMYTLCCR alpha,mouse) AP00184 P28794, MBP-1 (plant) RSGRGECRRQCLRRHEGQPWETQEC 727MRRCRRRG AP00050 P29002, Bombinin-like GIGASILSAGKSALKGLAKGLAEHFAN 728peptide 1 (BLP-1, toad) AP00051 P29003, Bombinin-likeGIGSAILSAGKSALKGLAKGLAEHFAN 729 peptide 2 (BLP-2, toad) AP00052 P29004,Bombinin-like GIGAAILSAGKSALKGLAKGLAEHF 730 peptide 3 (BLP-3, XXA, toad)AP00053 P29005, Bombinin-like GIGAAILSAGKSIIKGLANGLAEHF 731 peptide 4(BLP-4, toad) AP00634 P29430, Pediocin PA-1/ KYYGNGVTCGKHSCSVDWGKATTCII732 AcH (PedPA1, class IIA NNGAMAWATGGHQGNHKC bacteriocin, bacteria)AP00204 P29559, Nisin Z ITSISLCTPGCKTGALMGCNMKTATC 733 (lantibiotic,class I NCSIHVSK bacteriocin, bacteria) AP00130 P29561, Cecropin CGWLKKLGKRIERIGQHTRDATIQGLGI 734 (insect, fly) AQQAANVAATAR AP00001P31107, GLWSKIKEVGKEAAKAAAKAAGKAA 735 ADENOREGULIN LGAVSEAV (DermaseptinB2, Dermaseptin-B2, DRS- B2, DRS B2, frog) AP00228 P31529, Sapecin BLTCEIDRSLCLLHCRLKGYLRAYCSQQ 736 (insect, flesh fly) KVCRCVQ AP00229P31530, Sapecin C ATCDLLSGIGVQHSACALHCVFRGNR 737 (insect, flesh fly)GGYCTGKGICVCRN AP00218 P32195, Protegrin 2 RGGRLCYCRRRFCICV 738 (PG-2,pig cathelicidin) AP00219 P32196, Protegrin 3 RGGGLCYCRRRFCVCVGR 739(PG-3, pig cathelicidin) AP00073 P32412, Brevinin-1EFLPLLAGLAANFLPKIFCKITRKC 740 (frog) AP00080 P32414, Esculentin-1GIFSKLGRKKIKNLLISGLKNVGKEVG 741 (frog) MDVVRTGIDIAGCKIKGEC AP00074P32423, Brevinin-1 FLPVLAGIAAKVVPALFCKITKKC 742 (frog) AP00075 P32424,Brevinin-2 GLLDSLKGFAATAGKGVLQSLLSTAS 743 (frog) CKLAKTC AP00175 P34084,Macaque DSHEERHHGRHGHHKYGRKFHEKHH 744 histatin (M-Histatin 1,SHRGYRSNYLYDN primate, monkey) AP00006 P35581, Apidaecin IAGNNRPVYIPQPRPPHPRI 745 (insect, honeybee) AP00007 P35581, Apidaecin IBGNNRPVYIPQPRPPHPRL 746 (insect, honeybee) AP00414 P36190, Ceratotoxin ASIGSALKKALPVAKKIGKIALPIAKAA 747 (insect, fly) LP AP00415 P36191,Ceratotoxin B SIGSAFKKALPVAKKIGKAALPIAKAA 748 (insect, fly) LP AP00172P36193, Drosocin GKPRPYSPRPTSHPRPIRV 749 (insect) AP00170 P37362,Pyrrhocoricin VDKGSYLPRPTPPRPIYNRN 750 (insect) AP00635 P38577,Mesentericin KYYGNGVHCTKSGCSVNWGEAASAG 751 Y105 (MesY105, classIHRLANGGNGFW IIA bacteriocin, bacteria) AP00636 P38579,AISYGNGVYCNKEKCWVNKAENKQA 752 Carnobacteriocin BM1 ITGIVIGGWASSLAGMGH(CnbBM1, PiscV1b, class IIA bacteriocin, bacteria) AP00209 P39080,Peptide PGQ GVLSNVIGYLKKLGTGALNAVLKQ 753 (frog) AP00513 P39084,Ranalexin FLGGLIKIVPAMICAVTKKC 754 (frog) AP00071 P40835, Brevinin-1EAFLPAIFRMAAKVVPTIICSITKKC 755 (frog) AP00072 P40836, Brevinin-1EBVIPFVASVAAEMQHVYCAASRKC 756 (frog) AP00076 P40837, Brevinin-2EAGILDTLKNLAISAAKGAAQGLVNKAS 757 (frog) CKLSGQC AP00077 P40838,Brevinin-2EB GILDTLKNLAKTAGKGALQGLVKMA 758 (frog) SCKLSGQC AP00078P40839, Brevinin-2EC GILLDKLKNFAKTAGKGVLQSLLNTA 759 (frog) SCKLSGQCAP00079 P40840, Brevinin-2ED GILDSLKNLAKNAGQILLNKASCKLSG 760 (frog) QCAP00081 P40843, Esculentin-1A GIFSKLAGKKIKNLLISGLKNVGKEVG 761 (frog)MDVVRTGIDIAGCKIKGEC AP00082 P40844, Esculentin-1BGIFSKLAGKKLKNLLISGLKNVGKEVG 762 (frog) MDVVRTGIDIAGCKIKGEC AP00083P40845, Esculentin-2A GILSLVKGVAKLAGKGLAKEGGKFGL 763 (frog) ELIACKIAKQCAP00084 P40846, Esculentin-2B GIFSLVKGAAKLAGKGLAKEGGKFGL 764(ES2B_RANES, frog) ELIACKIAKQC AP00299 P46156, ChickenGRKSDCFRKSGFCAFLKCPSLTLISGK 765 gallinacin 1 (Gal 1, CSRFYLCCKRIW avianbeta-defensin, bird) AP00300 P46157, Gallinacin 1GRKSDCFRKNGFCAFLKCPYLTLISGK 766 alpha (avian beta- CSRFHLCCKRIWdefensin, Bird), AP00298 P46158, Chicken LFCKGGSCHFGGCPSHLIKVGSCFGFR 767gallinacin 2 (Gal 2, SCCKWPWNA avian beta-defensin, bird) AP00037P46160, Beta-defensin 2 VRNHVTCRINRGFCVPIRCPGRTRQIG 768 (cow)TCFGPRIKCCRSW AP00038 P46161, Beta-defensin 3QGVRNHVTCRINRGFCVPIRCPGRTRQ 769 (cow) IGTCFGPRIKCCRSW AP00039 P46162,Beta-defensin 4 QRVRNPQSCRWNMGVCIPFLCRVGM 770 (cow) RQIGTCFGPRVPCCRRAP00040 P46163, Beta-defensin 5 QVVRNPQSCRWNMGVCIPISCPGNMR 771 (cow)QIGTCFGPRVPCCRRW AP00041 P46164, Beta-defensin 6QGVRNHVTCRIYGGFCVPIRCPGRTR 772 (cow) QIGTCFGRPVKCCRRW AP00042 P46165,Beta-defensin 7 QGVRNFVTCRINRGFCVPIRCPGHRRQ 773 (cow) IGTCLGPRIKCCRAP00043 P46166, Beta-defensin 8 VRNFVTCRINRGFCVPIRCPGHRRQIG 774 (cow)TCLGPQIKCCR AP00044 P46167, Beta-defensin 9 QGVRNFVTCRINRGFCVPIRCPGHRRQ775 (cow) IGTCLAPQIKCCR AP00045 P46168, Beta-defensinQGVRSYLSCWGNRGICLLNRCPGRMR 776 10 (cow) QIGTCLAPRVKCCR AP00046 P46169,Beta-defensin GPLSCRRNGGVCIPIRCPGPMRQIGTC 777 11 (cow) FGRPVKCCRSWAP00048 P46171, Bovine beta- SGISGPLSCGRNGGVCIPIRCPVPMRQI 778 defensin13 (cow) GTCFGRPVKCCRSW AP00350 P48821, EnbocinPWNIFKEIERAVARTRDAVISAGPAVR 779 (insect, moth) TVAAATSVAS AP00173P49112, GNCP-2 RCICTTRTCRFPYRRLGTCLFQNRVYT 780 (Guinea pig neutrophilFCC cationic peptide 2) AP00369 P49930, PMAP-23 RIIDLLWRVRRPQKPKFVTVWVR781 (PMAP23, pig cathelicidin) AP00370 P49931, PMAP-36VGRFRRLRKKTRKRLKKIGKVLKWIP 782 (PMAP36, pig PIVGSIPLGCG cathelicidin)AP00371 P49932, PMAP-37 GLLSRLRDFLSDRGRRLGEKIERIGQKI 783 (PMAP37, pigKDLSEFFQS cathelicidin) AP00220 P49933, Protegrin 4 RGGRLCYCRGWICFCVGR784 (PG-4, pig cathelicidin) AP00221 P49934, Protegrin 5RGGRLCYCRPRFCVCVGR 785 (PG-5, pig cathelicidin) AP00346 P50720,Hyphancin IIID RWKIFKKIERVGQNVRDGIIKAGPAIQ 786 (Fall webworm, insect)VLGTAKAL AP00347 P50721, Hyphancin IIIE RWKFFKKIERVGQNVRDGLIKAGPAI 787(Fall webworm, insect) QVLGAAKAL AP00348 P50722, Hyphancin IIIFRWKVFKKIEKVGRNIRDGVIKAGPAI 788 (Fall webworm, insect) AVVGQAKAL AP00349P50723, Hyphancin IIIG RWKVFKKIEKVGRHIRDGVIKAGPAIT 789 (Fall webworm,insect) VVGQATAL AP00281 P51473, mCRAMP GLLRKGGEKIGEKLKKIGQKIKNFFQK 790(mouse cathelicidin; LVPQPEQ derivatives: CRAMP 18) AP00366 P54228,BMAP-27 GRFKRFRKKFKKLFKKLSPVIPLLHLG 791 (BMAP27, cow cathelicidin, ZZHs,derivatives BMAP-18 and BMAP-15) AP00367 P54229, BMAP-28GGLRSLGRKILRAWKKYGPIIVPIIRIG 792 (BMAP28, cow cathelicidin) AP00450P54230, Cyclic RICRIIFLRVCR 793 dodecapeptide (sheep cathelicidin)AP00359 P54684, Lebocin 1/2 DLRFLYPRGKLPVPTPPPFNPKPIYIDM 794 (insect,silk moth) GNRY AP00360 P55796, Lebocin 3 DLRFLYPRGKLPVPTLPPFNPKPIYIDM795 (insect, silk moth) GNRY AP00307 P55897, Buforin I (toad)AGRGKQGGKVRAKAKTRSSRAGLQF 796 PVGRVHRLLRKGNY AP00308 P55897, Buforin IITRSSRAGLQFPVGRVHRLLRK 797 (toad) AP00240 P56226, Caerin 1.1GLLSVLGSVAKHVLPHVVPVIAEHL 798 (frog, ZZHa) AP00241 P56227, Caerin 1.2GLLGVLGSVAKHVLPHVVPVIAEHL 799 (frog) AP00242 P56228, Caerin 1.3GLLSVLGSVAQHVLPHVVPVIAEHL 800 (frog) AP00243 P56229, Caerin 1.4GLLSSLSSVAKHVLPHVVPVIAEHL 801 (frog) AP00244 P56230, Caerin 1.5GLLSVLGSVVKHVIPHVVPVIAEHL 802 (frog) AP00245 P56231, Caerin 1.6GLFSVLGAVAKHVLPHVVPVIAEK 803 (frog) AP00246 P56232, Caerin 1.7GLFKVLGSVAKHLLPHVAPVIAEK 804 (frog) AP00249 P56233, Caerin 2.1GLVSSIGRALGGLLADVVKSKGQPA 805 (frog) AP00250 P56234, Caerin 2.2GLVSSIGRALGGLLADVVKSKEQPA 806 (frog) AP00251 P56236, Caerin 2.4GLVSSIGKALGGLLADVVKTKEQPA 807 (frog) AP00252 P56236, Caerin 2.5GLVSSIGRALGGLLADVVKSKEQPA 808 (frog) AP00253 P56238, Caerin 3.1GLWQKIKDKASELVSGIVEGVK 809 (frog) AP00254 P56238, Caerin 3.2GLWEKIKEKASELVSGIVEGVK 810 (frog) AP00255 P56240, Caerin 3.3GLWEKIKEKANELVSGIVEGVK 811 (frog) AP00256 P56241, Caerin 3.4GLWEKIREKANELVSGIVEGVK 812 (frog) AP00257 P56242, Caerin 4.1GLWQKIKSAAGDLASGIVEGIKS 813 (frog) AP00258 P56243, Caerin 4.2GLWQKIKSAAGDLASGIVEAIKS 814 (frog) AP00259 P56244, Caerin 4.3GLWQKIKNAAGDLASGIVEGIKS 815 (frog) AP00434 P56249, Frenatin 3GLMSVLGHAVGNVLGGLFKS 816 (frog) AP00272 P56386, Murine beta-DQYKCLQHGGFCLRSSCPSNTKLQGT 817 defensin 1 (mBD-1, CKPDKPNCCKS mouse)AP00368 P56425, BMAP-34 GLFRRLRDSIRRGQQKILEKARRIGERI 818 (BMAP34, cowKDIFRG cathelicidin) AP00273 P56685, ButhininSIVPIRCRSNRDCRRFCGFRGGRCTYA 819 (Sahara scorpion) RQCLCGY AP00282P56872, SIPCGESCVFIPCTVTALLGCSCKSKVC 820 Cyclopsychotride A YKN (CPT,plant cyclotides, XXC) AP00094 P56917, Temporin A FLPLIGRVLSGIL 821(XXA, frog) AP00096 P56918, Temporin C LLPILGNLLNGLL 822 (XXA, frog)AP00097 P56920, Temporin E VLPIIGNLLNSLL 823 (XXA, frog) AP00098 P56921,Temporin F FLPLIGKVLSGIL 824 (XXA, frog) AP00100 P56923, Temporin KLLPNLLKSLL 825 (XXA, frog) AP00295 P56928, eNAP-2 (horse)EVERKHPLGGSRPGRCPTVPPGTFGHC 826 ACLCTGDASEPKGQKCCSN AP00101 P57104,Temporin L FVQWFSKFLGRIL 827 (XXA, frog) AP00095 P79874, Temporin BLLPIVGNLLKSLL 828 (XXA, frog) AP00099 P79875, Temporin G FFPVIGRILNGIL829 (XXA, frog) AP00413 P80032, Coleoptericin SLQGGAPNFPQPSQQNGGWQVSPDLG830 (insect) RDDKGNTRGQIEIQNKGKDHDFNAG WGKVIRGPNKAKPTWHVGGTYRR AP00396P80054, PR-39 (PR39, RRRPRPPYLPRPRPPPFFPPRLPPRIPPG 831 pig cathelicidin)FPPRFPPRFP AP00182 P80154, Insect defensin GFGCPLDQMQCHRHCQTITGRSGGYC832 SGPLKLTCTCYR AP00444 P80223, CorticostatinGICACRRRFCLNFEQFSGYCRVNGAR 833 VI (CS-VI) (animal YVRCCSRR defensin,rabbit) AP00208 P80230, Peptide 3910 RADTQTYQPYNKDWIKEKIYVLLRRQ 834(pig) AQQAGK AP00157 P80277, Dermaseptin- ALWKTMLKKLGTMALHAGKAALGA 835S1 (Dermaseptin S1, AADTISQGTQ DRS S1, DRS-S1, frog) AP00158 P80278,Dermaseptin- ALWFTMLKKLGTMALHAGKAALGA 836 S2 (Dermaseptin S2, AANTISQGTQDRS S2, DRS-S2, frog) AP00159 P80279, Dermaseptin-ALWKNMLKGIGKLAGKAALGAVKKL 837 S3 (Dermaseptin S3, VGAES DRS S3, DRS-S3,frog) AP00160 P80280, Dermaseptin- ALWMTLLKKVLKAAAKALNAVLVG 838 S4(Dermaseptin S4, ANA DRS S4, DRS-S4, frog) AP00161 P80281, Dermaseptin-GLWSKIKTAGKSVAKAAAKAAVKAV 839 S5 (Dermaseptin S5, TNAV DRS S5, DRS-S5,frog) AP00293 P80282, Dermaseptin- AMWKDVLKKIGTVALHAGKAALGA 840 B1(DRS-B1, DRS B1, VADTISQ frog) AP00264 P80389, ChickenGRKSDCFRKSGFCAFLKCPSLTLISGK 841 Heterophil Peptide 1 CSRFYLCCKRIR (CHP1,bird, animal) AP00265 P80390, Chicken GRKSDCFRKNGFCAFLKCPYLTLISGL 842Heterophil Peptide 2 CSFHLC (CHP2, bird, animal) AP00266 P80391, TurkeyGKREKCLRRNGFCAFLKCPTLSVISGT 843 Heterophil Peptide 1 CSRFQVCC (THP1,turkey) AP00267 P80392, Turkey LFCKRGTCHFGRCPSHLIKVGSCFGFR 844Heterophil Peptide 2 SCCKWPWDA (THP2, bird, anaimal) AP00269 P80393,Turkey LSCKRGTCHFGRCPSHLIKGSCSGG 845 Heterophil Peptide 3 (THP3, bird,animal) AP00085 P80395, Gaegurin-1 SLFSLIKAGAKFLGKNLLKQGACYAA 846(Gaegurin 1, frog) CKASKQC AP00086 P80396, Gaegurin-2GIMSIVKDVAKNAAKEAAKGALSTLS 847 (Gaegurin 2, frog) CKLAKTC AP00087P80397, Gaegurin-3 GIMSIVKDVAKTAAKEAAKGALSTLS 848 (Gaegurin 3, frog)CKLAKTC AP00089 P80399, Gaegurin-5 FLGALFKVASKVLPSVFCAITKKC 849(Gaegurin 5, frog) AP00090 P80400, Gaegurin-6 FLPLLAGLAANFLPTIICKISYKC850 (Gaegurin 6, frog) AP00362 P80408, Metalnikowin I VDKPDYRPRPRPPNM851 (insect) AP00363 P80409, Metalnikowin VDKPDYRPRPWPRPN 852 IIA(insect) AP00364 P80410, Metalnikowin VDKPDYRPRPWPRNMI 853 IIB (insect)AP00365 P80411, Metalnikowin VDKPDYRPRPWPRPNM 854 III (insect) AP00632P80569, Piscicolin 126/ KYYGNGVSCNKNGCTVDWSKAIGIIG 855 Piscicocin ViaNNAAANLTTGGAAGWNKG (PiscV1a, Pisc126, class IIA bacteriocin, bacteria)AP01003 P80666, Mutacin B- FKSWSFCTPGCAKTGSFNSYCC 856 Ny266 (bacteria)AP00276 P80710, Clavanin A VFQFLGKIIHHVGNFVHGFSHVF 857 (urochordates,sea squirts, and sea pork, tunicate) AP00277 P80711, Clavanin BVFQFLGRIIHHVGNFVHGFSHVF 858 (Sea squirt, tunicate) AP00278 P80712,Clavanin C VFHLLGKIIHHVGNFVYGFSHVF 859 (Sea squirt, tunicate) AP00279P80713, Clavanin D AFKLLGRIIHHVGNFVYGFSHVF 860 (Sea squirt, tunicate)AP00280 P80713, Clavanin D LFKLLGKIIHHVGNFVHGFSHVF 861 (Sea squirt,tunicate) AP00294 P80930, eNAP-1 (horse) DVQCGEGHFCHDQTCCRASQGGACC 862PYSQGVCCADQRHCCPVGF AP00400 P80952, Skin peptideYPPKPESPGEDASPEEMNKYLTALRH 863 tyrosine-tyrosine (skin- YINLVTRQRY PYY,SPYY, frog) AP00091 P80954, Rugosin A GLLNTFKDWAISIAKGAGKGVLTTLS 864(frog) CKLDKSC AP00092 P80955, Rugosin B SLFSLIKAGAKFLGKNLLKQGAQYAA 865(frog) CKVSKEC AP00093 P80956, Rugosin C GILDSFKQFAKGVGKDLIKGAAQGVL 866(frog) STMSCKLAKTC AP00392 P81056, Penaeidin-1YRGGYTGPIPRPPPIGRPPLRLVVCAC 867 (shrimp, Crustacea)YRLSVSDARNCCIKFGSCCHLVK AP00393 P81057, Penaeidin-2aYRGGYTGPIPRPPPIGRPPFRPVCNACY 868 (shrimp, Crustacea)RLSVSDARNCCIKFGSCCHLVK AP00394 P81058, Penaeidin-3aQVYKGGYTRPIPRPPPFVRPLPGGPIGP 869 (shrimp, Crustacea)YNGCPVSCRGISFSQARSCCSRLGRCC HVGKGYS AP00247 P81251, Caerin 1.8GLFKVLGSVAKHLLPHVVPVIAEK 870 (frog) AP00248 P81252, Caerin 1.9GLFGVLGSIAKHVLPHVVPVIAEK 871 (frog, ZZHa) AP00126 P81417, Cecropin AGGLKKLGKKLEGVGKRVFKASEKAL 872 (insect, mosquito) PVAVGIKALG AP00169P81437, Formaecin 2 GRPNPVNTKPTPYPRL 873 (insect, ants) AP00168 P81438,Formaecin 1 GRPNPVNNKPTPHPRL 874 (insect, ants) AP00296 P81456,Fabatin-1 LLGRCKVKSNRFHGPCLTDTHCSTVC 875 (plant defensin)RGEGYKGGDCHGLRRRCMCLC AP00297 P81457, Fabatin-2LLGRCKVKSNRFNGPCLTDTHCSTVC 876 (plant defensin) RGEGYKGGDCHGLRRRCMCLCAP01215 P81463, European FVPYNPPRPYQSKPFPSFPGHGPFNPKI 877 bumblebeeabaecin QWPYPLPNPGH (insect) AP01214 P81464, Apidaecin GNRPVYIPPPRPPHPRL878 (insect) AP00440 P81465, defensin VTCFCRRRGCASRERHIGYCRFGNTIY 879HANP-1 (hamster) RLCCRR AP00441 P81466, defensinCFCKRPVCDSGETQIGYCRLGNTFYRL 880 HANP-2 (hamster) CCRQ AP00442 P81467,defensin VTCFCRRRGCASRERLIGYCRFGNTIY 881 HANP-3 (hamster) GLCCRR AP00439P81468, defensin VTCFCKRPVCDSGETQIGYCRLGNTF 882 HANP-4 (hamster) YRLCCRQAP00328 P81469, Styelin A (Sea GFGKAFHSVSNFAKKHKTA 883 squirt, tunicate,XXA) AP00329 P81470, Styelin B (Sea GFGPAFHSVSNFAKKHKTA 884 squirt,tunicate, XXA) AP00492 P81474, Misgurin (fish) RQRVEELSKFSKKGAAARRRK 885AP00165 P81485, Dermaseptin- ALWKNMLKGIGKLAGQAALGAVKTL 886 B3(Dermaseptin B3, VGAE DRS-B3, DRS B3, frog) AP00163 P81486, Dermaseptin-ALWKDILKNVGKAAGKAVLNTVTDM 887 B4 (Dermaseptin B4, VNQ DRS-B4, DRS B4,DRS-TR1, IRP, frog) AP00162 P81487, Dermaseptin-GLWNKIKEAASKAAGKAALGFVNEMV 888 B5 (Dermaseptin B5, DRS-B5, DRS B5, frog)AP00164 P81488, Dermaseptin- ALWKTIIKGAGKMIGSLAKNLLGSQA 889 B9(Dermaseptin B9, QPES DRS-B9, DRS DRG3, frog) AP00167 P81565, PhylloxinGWMSKIASGIGTFLSGMQQ 890 (phylloxin-B1, PLX-B1, XXA, frog) AP00291P81568, Defensin D5 MFFSSKKCKTVSKTFRGPCVRNAN 891 (So-D5) (plantdefensin) AP00290 P81569, Defensin D4 MFFSSKKCKTVSKTFRGPCVRNA 892(So-D4) (plant defensin) AP00289 P81570, Defensin D3GIFSSRKCKTVSKTFRGICTRNANC 893 (So-D3) (plant defensin) AP00288 P81572,Defensin D1 TCESPSHKFKGPCATNRNCES 894 (So-D1) (plant defensin) AP00292P81573, Defensin D7 GIFSSRKCKTPSKTFKGYCTRDSNCDT 895 (So-D7) (plantdefensin) SCRYEGYPAGD AP00270 P81591, Pn-AMP QQCGRQASGRLCGNRLCCSQWGYCG896 (PnAMP, plant STASYCGAGCQSQCRS defensin) AP00412 P81592, AcaloleptinA1 SLQPGAPNVNNKDQPWQVSPHISRDD 897 (insect) SGNTRTDINVQRHGENNDFEAGWSKVVRGPNKAKPTWHIGGTHRW AP00433 P81605, human SSLLEKGLDGAKKAVGGLGKLGKDA 898Dermcidin (DCD-1) VEDLESVGKGAVHDVKDVLDSV AP00332 P81612, Mytilin AGCASRCKAKCAGRRCKGWASASFRG 899 (Blue mussel) RCYCKCFRC AP00333 P81613,Mytilin B SCASRCKGHCRARRCGYYVSVLYRG 900 (Blue mussel) RCYCKCLRC AP00334P81613, Moronecidin FFHHIFRGIVHVGKTIHKLVTG 901 (fish) AP00351 P81835,Citropin 1.1 GLFDVIKKVASVIGGL 902 (amphibian, frog) AP00352 P81840,Citropin 1.2 GLFDIIKKVASVVGGL 903 (amphibian, frog) AP00353 P81846,Citropin 1.3 GLFDIIKKVASVIGGL 904 (amphibian, frog) AP00338 P81903,Histone H2B- PDPAKTAPKKGSKKAVTKA 905 1(HLP-1) (fish) AP00271 P82018,ChBac5 (Goat RFRPPIRRPPIRPPFNPPFRPPVRPPFRPP 906 cathelicidin)FRPPFRPPIGPFP AP00316 P82027, Uperin 2.1 GIVDFAKKVVGGIRNALGI 907(amphibian, toad) AP00317 P82028, Uperin 2.2 GFVDLAKKVVGGIRNALGI 908(amphibian, toad) AP00318 P82029, Uperin 2.3 GFFDLAKKVVGGIRNALGI 909(amphibian, toad) AP00319 P82030, Uperin 2.4 GILDFAKTVVGGIRNALGI 910(amphibian, toad) AP00320 P82031, Uperin 2.5 GIVDFAKGVLGKIKNVLGI 911(amphibian, toad) AP00323 P82032, Uperin 3.1 GVLDAFRKIATVVKNVV 912(amphibian, toad) AP00326 P82035, Uperin 4.1 GVGSFIHKVVSAIKNVA 913(amphibian, toad) AP00321 P82039, Uperin 2.7 GIIDIAKKLVGGIRNVLGI 914(amphibian, toad) AP00322 P82040, Uperin 2.8 GILDVAKTLVGKLRNVLGI 915(amphibian, toad) AP00324 P82042, Uperin 3.5 GVGDLIRKAVSVIKNIV 916(amphibian, toad) AP00325 P82042, Uperin 3.6 GVIDAAKKVVNVLKNLP 917(amphibian, toad) AP00327 P82050, Uperin 7.1 GWFDVVKHIASAV 918(amphibian, frog) AP00260 P82066, Maculatin 1.1 GLFVGVLAKVAAHVVPAIAEHF919 (XXA, frog, ZZHa) AP00261 P82067, Maculatin 1.2GLFVGLAKVAAHNNPAIAEHFQA 920 (XXA, frog) AP00262 P82068, Maculatin 2.1GFVDFLKKVAGTIANVVT 921 (frog) AP00263 P82069, Maculatin 3.1GLLQTIKEKLESLESLAKGIVSGIQA 922 (frog) AP00345 P82104, Caerin 1.10GLLSVLGSVAKHVLPHVVPVIAEKL 923 (frog) AP00456 P82232, Brevinin-1TVNPIILGVLPKFVCLITKKC 924 (frog) AP00459 P82233, Brevinin-1TAFITLLLRKFICSITKKC 925 (frog) AP00457 P82234, Brevinin-2TCGLWETIKNFGKKFTLNILHKLKCKIGG 926 (frog) GC AP00458 P82235, Brevinin-2TDGLWETIKNFGKKFTLNILHNLKCKIGG 927 (frog) GC AP00397 P82238, Salmocidin 2ASGFVLKGYTKTSQ 928 (fish, trout) AP00398 P82239, Salmocidin 2BAGFVLKGYTKTSQ 929 (fish, trout) AP00055 P82282, Bombinin H1IIGPVLGMVGSALGGLLKKI 930 (frog) AP00056 P82284, Bombinin H4LIGPVLGLVGSALGGLLKKI 931 (frog, XXA, XXD) AP00057 P82285, Bombinin H5IIGPVLGLVGSALGGLLKKI 932 (frog, XXD) AP00419 P82286, Bombinin-likeGIGASILSAGKSALKGFAKGLAEHFAN 933 peptides 2 (amphibian, toad) AP00137P82293, Cryptdin-1 LRDLVCYCRTRGCKRRERMNGTCRK 934 (Crp1, animal defensin,GHLMYTLCCR alpha, mouse) AP00443 P82317, defensinACYCRIPACLAGERRYGTCFYMGRV 935 RMAD-2 (monkey) WAFCC AP00012 P82386,Aurein 1.1 GLFDIIKKIAESI 936 (amphibian, frog) AP00014 P82388, Aurein2.1 GLLDIVKKVVGAFGSL 937 (amphibian, frog) AP00015 P82389, Aurein 2.2GLFDIVKKVVGALGSL 938 (amphibian, frog) AP00016 P82390, Aurein 2.3GLFDIVKKVVGAIGSL 939 (XXA, amphibian, frog) AP00017 P82391, Aurein 2.4GLFDIVKKVVGTIAGL 940 (XXA, amphibian, frog) AP00018 P82392, Aurein 2.5GLFDIVKKVVGAFGSL 941 (XXA, amphibian, frog) AP00019 P82393, Aurein 2.6GLFDIAKKVIGVIGSL 942 (XXA, amphibian, frog) AP00020 P82394, Aurein 3.1GLFDIVKKIAGHIAGSI 943 (XXA, amphibian, frog) AP00021 P82395, Aurein 3.2GLFDIVKKIAGHIASSI 944 (XXA, amphibian, frog) AP00022 P82396, Aurein 3.3GLFDIVKKIAGHIVSSI 945 (XXA, amphibian, frog) AP00376 P82414, PonericinG1 GWKDWAKKAGGWLKKKGPGMAKA 946 (ants) ALKAAMQ AP00377 P82415, PonericinG2 GWKDWLKKGKEWLKAKGPGIVKAA 947 (ants) LQAATQ AP00378 P82416, PonericinG3 GWKDWLNKGKEWLKKKGPGIMKAA 948 (ants) LKAATQ AP00379 P82417, PonericinG4 DFKDWMKTAGEWLKKKGPGILKAA 949 (ants) MAAAT AP00380 P82418, PonericinG5 GLKDWVKIAGGWLKKKGPGILKAAM 950 (ants) AAATQ AP00381 P82419, PonericinG6 GLVDVLGKVGGLIKKLLP 951 (ants) AP00382 P82420, Ponericin G7GLVDVLGKVGGLIKKLLPG 952 (ants) AP00383 P82421, Ponericin L1LLKELWTKMKGAGKAVLGKIKGLL 953 (ants) AP00384 P82422, Ponericin L2LLKELWTKIKGAGKAVLGKIKGLL 954 (ants) AP00386 P82423, Ponericin W1WLGSALKIGAKLLPSVVGLFKKKKQ 955 (ants) AP00387 P82424, Ponericin W2WLGSALKIGAKLLPSVVGLFQKKKK 956 (ants) AP00388 P82425, Ponericin W3GIWGTLAKIGIKAVPRVISMLKKKKQ 957 (ants) AP00389 P82426, Ponericin W4GIWGTALKWGVKLLPKLVGMAQTKKQ 958 (ants) AP00390 P82427, Ponericin W5FWGALIKGAAKLIPSVVGLFKKKQ 959 (ants) AP00391 P82428, Ponericin W6FIGTALGIASAIPAIVKLFK 960 (ants) AP00303 P82651, Tigerinin-1 FCTMIPIPRCY961 (frog) AP00304 P82652, Tigerinin-2 RVCFAIPLPICH 962 (frog) AP00305P82653, Tigerinin-3 RVCYAIPLPICY 963 (frog) AP00301 P82656, HadrurinGILDTIKSIASKVWNSKTVQDLKRKGI 964 (scorpion) NWVANKLGVSPQAA AP00113P82740, GLLSGLKKVGKHVAKNVAVSLMDSL 965 RANATUERIN 1T KCKISGDC (frog)AP00114 P82741, SMLSVLKNLGKVGLGFVACKINKQC 966 RANATUERIN 1(Ranatuerin-1, frog) AP00115 P82742, GLFLDTLKGAAKDVAGKLEGLKCKIT 967RANATUERIN 2 GCKLP (Ranatuerin-2, frog) AP00116 P82780,GFLDIINKLGKTFAGHMLDKIKCTIGT 968 RANATUERIN 3 CPPSP (Ranatuerin-3, frog)AP00117 P82819, FLPFIARLAAKVFPSIICSVTKKC 969 RANATUERIN 4 (Ranatuerin-4,frog) AP00405 P82821, FISAIASMLGKFL 970 RANATUERIN 6 (frog) AP00406P82822, FLSAIASMLGKFL 971 RANATUERIN 7 (frog) AP00407 P82823,FISAIASFLGKFL 972 RANATUERIN 8 (frog) AP00408 P82824, FLFPLITSFLSKVL 973RANATUERIN 9 (frog) AP00461 P82825, Brevinin-1LAFLPMLAGLAASMVPKLVCLITKKC 974 (frog) AP00462 P82826, Brevinin-1LBFLPMLAGLAASMVPKFVCLITKKC 975 (frog) AP00118 P82828,GILDSFKGVAKGVAKDLAGKLLDKLK 976 RANATUERIN 2La CKITGC (Ranatuerin-2La,frog) AP00119 P82829, GILSSIKGVAKGVAKNVAAQLLDTLK 977 RANATUERIN 2LbCKITGC (Ranatuerin-2Lb, frog) AP00109 P82830, Temporin-1La VLPLISMALGKLL978 (Temporin 1La, frog) AP00110 P82831, Temporin-1Lb NFLGTLINLAKKIM 979(Temporin 1Lb, frog) AP00111 P82832, Temporin-1Lc FLPILINLIHKGLL 980(Temporin 1Lc, frog) AP00463 P82833, Brevinin-1BAFLPFIAGMAAKFLPKIFCAISKKC 981 (frog) AP00464 P82834, Brevinin-1BBFLPAIAGMAAKFLPKIFCAISKKC 982 (frog) AP00465 P82835, Brevinin-1BCFLPFIAGVAAKFLPKIFCAISKKC 983 (frog) AP00466 P82836, Brevinin-1BDFLPAIAGVAAKFLPKIFCAISKKC 984 (frog) AP00467 P82837, Brevinin-1BEFLPAIVGAAAKFLPKIFCVISKKC 985 (frog) AP00468 P82838, Brevinin-1BFFLPFIAGMAANFLPKIFCAISKKC 986 (frog) AP00120 P82840,GLLDTIKGVAKTVAASMLDKLKCKIS 987 RANATUERIN 2B GC (Ranatuerin-2B, frog)AP00469 P82841, Brevinin-1PA FLPIIAGVAAKVFPKIFCAISKKC 988 (frog) AP00460P82842, Brevinin-1PB FLPIIAGIAAKVFPKIFCAISKKC 989 (frog) AP00470 P82843,Brevinin-1PC FLPIIASVAAKVFSKIFCAISKKC 990 (frog) AP00471 P82844,Brevinin-1PD FLPIIASVAANVFSKIFCAISKKC 991 (frog) AP00472 P82845,Brevinin-1PE FLPIIASVAAKVFPKIFCAISKKC 992 (frog) AP00121 P82847,GLMDTVKNVAKNLAGHMLDKLKCKI 993 RANATUERIN 2P TGC (Ranatuerin-2P, frog)AP00112 P82848, Temporin-1P FLPIVGKLLSGLL 994 (Temporin 1P, frog)AP00452 P82871, Brevinin-1SY FLPVVAGLAAKVLPSIICAVTKKC 995 (frog) AP00122P82875, Ranatuerin-1C SMLSVLKNLGKVGLGLVACKINKQC 996 (Ranatuerin 1C,frog) AP00514 P82876, Ranalexin-1Ca FLGGLMKAFPALICAVTKKC 997 (frog)AP00515 P82877, Ranalexin-1Cb FLGGLMKAFPAIICAVTKKC 998 (frog) AP00124P82878, Ranatuerin-2Ca GLFLDTLKGAAKDVAGKLLEGLKCKI 999 (Ranatuerin 2Ca,frog) AGCKP AP00123 P82879, Ranatuerin- GLFLDTLKGLAGKLLQGLKCIKAGCKP 10002Cb (Ranatuerin 2Cb, frog) AP00104 P82880, Temporin-1Ca FLPFLAKILTGVL1001 (Temporin 1Ca, frog) AP00105 P82881, Temporin-1Cb FLPLFASLIGKLL1002 (Temporin 1Cb, frog) AP00106 P82882, Temporin-1Cc FLPFLASLLTKVL1003 (Temporin 1Cc, frog) AP00107 P82883, Temporin-1Cd FLPFLASLLSKVL1004 (Temporin 1Cd, frog) AP00108 P82884, Temporin-1Ce FLPFLATLLSKVL1005 (Temporin 1Ce, frog) AP00453 P82904, Brevinin-1SAFLPAIVGAAGQFLPKIFCAISKKC 1006 (frog) AP00454 P82905, Brevinin-1SBFLPAIVGAAGKFLPKIFCAISKKC 1007 (frog) AP00455 P82906, Brevinin-1SCFFPIVAGVAGQVLKKIYCTISKKC 1008 (frog) AP00996 P82907, LicheninISLEICAIFHDN 1009 (bacteria) AP00302 P82951, Hepcidin (fish)GCRFCCNCCPNMSGCGVCCRF 1010 AP00058 P83080, Maximin 1GIGTKILGGVKTALKGALKELASTYAN 1011 (toad) AP00059 P83081, Maximin 2GIGTKILGGVKTALKGALKELASTYVN 1012 (toad) AP00060 P83082, Maximin 3GIGGKILSGLKTALKGAAKELASTYLH 1013 (toad, ZZHa) AP00061 P83083, Maximin 4GIGGVLLSAGKAALKGLAKVLAEKYAN 1014 (toad) AP00062 P83084, Maximin 5SIGAKILGGVKTFFKGALKELASTYLQ 1015 (toad) AP00063 P83085, Maximin 6ILGPVISTIGGVLGGLLKNL 1016 (toad) AP00064 P83086, Maximin 7ILGPVLGLVGNALGGLIKNE 1017 (toad) AP00065 P83087, Maximin 8ILGPVLSLVGNALGGLLKNE 1018 (toad) AP00355 P83171, GinkbilobinANTAFVSSAHNTQKIPAGAPFNRNLR 1019 (Chinese plant) AMLADLRQNAAFAG AP00475P83188, Pseudin 1 GLNTLKKVFQGLHEAIKLINNHVQ 1020 (frog) AP00476 P83189,Pseudin 2 GLNALKKVFQGIHEAIKLINNHVQ 1021 (frog) AP00477 P83190, Pseudin 3GINTLKKVIQGLHEVIKLVSNHE 1022 (frog) AP00478 P83191, Pseudin 4GINTLKKVIQGLHEVIKLVSNHA 1023 (frog) AP00410 P83287, OncorhyncinSKGKKANKDVELARG 1024 III (fish) AP00357 P83305, Japonicin-1FFPIGVFCKIFKTC 1025 (amphibian, frog) AP00358 P83306, Japonicin-2FGLPMLSILPKALCILLKRKC 1026 (amphibian, frog) AP00385 P83312,Parabutoporin FKLGSFLKKAWKSKLAKKLRAKGKE 1027 (scorpion)MLKDYAKGLLEGGSEEVPGQ AP00374 P83313, Opistoporin 1GKVWDWIKSTAKKLWNSEPVKELKN 1028 (scorpion) TALNAAKNLVAEKIGATPS AP00375P83314, Opistoporin 2 GKVWDWIKSTAKKLWNSEPVKELKN 1029 (scorpion)TALNAAKNFVAEKIGATPS AP00336 P83327, Histone H2A AERVGAGAPVYL 1030 (fish)AP00335 P83338, Histone H6- PKRKSATKGDEPA 1031 like protein (fish)AP00411 P83374, Oncorhyncin II KAVAAKKSPKKAKKPAT 1032 (fish) AP00999P83375, Serracin-P 43 kDa DYHHGVRVL 1033 subunit (bacteria) AP00284P83376, Dolabellanin SHQDCYEALHKCMASHSKPFSCSMKF 1034 B2 (sea hare)HMCLQQQ AP00998 P83378, Serracin-P 23 kDa ALPKKLKYLNLFNDGFNYMGVV 1035subunit (bacteriocin, bacteria) AP00129 P83403, CecropinGWLKKIGKKIERVGQNTRDATVKGLE 1036 (insect, moth) VAQQAANVAATVR AP00127P83413, Cecropin A RWKVFKKIEKVGRNIRDGVIKAAPAIE 1037 (insect, moth)VLGQAKAL AP00372 P83416, Virescein GKIPIGAIKKAGKAIGKGLRAVNIAST 1038(insect) AHDVYTFFKPKKRH AP00356 P83427, Heliocin QRFIHPTYRPPPQPRRPVIMRA1039 (insect) AP00409 P83428, Locustin ATTGCSCPQCIIFDPICASSYKNGRRGF 1040(insect) SSGCHMRCYNRCHGTDYFQISKGSKCI AP00339 P83545, Chrysophsin-1FFGWLIKGAIHAGKAIHGLIHRRRH 1041 (Red sea bream, madai) AP00340 P83546,Chrysophsin-2 FFGWLIRGAIHAGKAIHGLIHRRRH 1042 (Red sea bream, madai)AP00341 P83547, Chrysophsin-3 FIGLLISAGKAIHDLIRRRH 1043 (Red sea bream,madai) AP01004 P84763, Thuricin-S DWTAWSALVAAACSVELL 1044 (bacteria)AP00553 P84868, Sesquin (plant, KTCENLADTY 1045 ZZHp) AP00132 Q06589,Cecropin 1 GWLKKIGKKIERVGQHTRDATIQTIAV 1046 (insect, fly) AQQAANVAATARAP00135 Q06590, Cecropin 2 GWLKKIGKKIERVGQHTRDATIQTIGV 1047 (insect fly)AQQAANVAATLK AP00416 Q17313, Ceratotoxin C SLGGVISGAKKVAKVAIPIGKAVLPVV1048 (insect, fly) AKLVG AP00171 Q24395, MetchnikowinHRHQGPIFDTRPSPFNPNQPRPGPIY 1049 (insect) AP00354 Q27023, Tenecin 1VTCDILSVEAKGVKLNDAACAAHCLF 1050 (insect) RGRSGGYCNGKRVCVCR AP00401Q28880, Lingual GFTQGVRNSQSCRRNKGICVPIRCPGS 1051 antimicrobial peptideMRQIGTCLGAQVKCCRRK (LAP, beta defensin, cow) AP00224 Q62713, RatNP-3(rat) CSCRTSSCRFGERLSGACRLNGRIYRL 1052 CC AP00225 Q62714, RatNP-4 (rat)ACYCRIGACVSGERLTGACGLNGRIY 1053 RLCCR AP00223 Q62715, RatNP-2 (rat)VTCYCRSTRCGFRERLSGACGYRGRI 1054 YRLCCR AP00222 Q62716, RatNP-1 (rat)VTCYCRRTRCGFRERLSGACGYRGRI 1055 YRLCCR AP00174 Q64365, GNCP-1RRCICTTRTCRFPYRRLGTCIFQNRVY 1056 (Guinea pig neutrophil TFCC cationicpeptide 1) AP00311 Q90W78, Galensin CYSAAKYPGFQEFINRKYKSSRF 1057 (frog)AP00395 Q95NT0, Penaeidin-4a HSSGYTRPLPKPSRPIFIRPIGCDVCYGI 1058 (shrimp,Crustacea) PSSTARLCCFRYGDCCHR AP00423 Q962B0, Penaeidin-3nQGYKGPYTRPILRPYVRPVVSYNACT 1059 (shrimp, Crustacea)LSCRGITTTQARSCSTRLGRCCHVAKG YS AP00422 Q962B1, Penaeidin-3mQGCKGPYTRPILRPYVRPVVSYNACT 1060 (shrimp, Crustacea)LSCRGITTTQARSCCTRLGRCCHVAK GYS AP00421 Q963C3, Penaeidin-4CYSSGYTRPLPKPSRPIFIRPIGCDVCYGI 1061 (shrimp, Crustacea)PSSTARLCCFRYGDCCHR AP00210 Q99134, PGLa (African GMASKAGAIAGKIAKVALKAL1062 clawed frog, XXA) AP00054 Q9DET7, Bombinin-GIGGALLSAGKSALKGLAKGLAEHFAN 1063 like peptide 7 (BLP-7, toad) AP00315Q9PT75, Dermatoxin SLGSFLKGVGTTLASVGKVVSDQFGK 1064 (Two-colored leaffrog) LLQAGQ AP00133 Q9Y0Y0, Cecropin B GGLKKLGKKLEGVGKRVFKASEKAL 1065(insect, mosquito) PVLTGYKAIG AP00004 Ref, Ct-AMP1NLCERASLTWTGNCGNTGHCDTQCR 1066 (CtAMP1, C. ternatea-NWESAKHGACHKRGNWKCFCYFDC antimicrobial peptide 1, plant defensin)AP00027 Ref, hexapeptide RRWQWR 1067 (synthetic) AP00529 Ref,Lantibiotic Ericin S WKSESVCTPGCVTGVLQTCFLQTITC 1068 (bacteria) NCHISKAP00306 Ref, Tigerinin-4 (frog) RVCYAIPLPIC 1069 AP00309 Ref, HumanKS-27 KSKEKIGKEFKRIVQRIKDFLRNLVPR 1070 (KS27 from LL-37) AP00344 Ref,Apidaecin II GNNRPIYIPQPRPPHPRL 1071 (honeybee, insect) AP00424 Ref, XT1(frog) GFLGPLLKLAAKGVAKVIPHLIPSRQQ 1072 AP00425 Ref, XT 2 (frog)GCWSTVLGGLKKFAKGGLEAIVNPK 1073 AP00426 Ref, XT 4 (frog)GVFLDALKKFAKGGMNAVLNPK 1074 AP00427 Ref, XT 7 (frog) GLLGPLLKIAAKVGSNLL1075 AP00431 Ref, human LLP 1 RVIEVVQGACRAIRHIPRRIRQGLERIL 1076 AP00432Ref, human LLP RIAGYGLRGLAVIIRICIRGLNLIFEIIR 1077 AP00447 Ref, Anoplin(insect) GLLKRIKTLL 1078 AP00474 Ref, Piscidin 3 (fish)FIHHIFRGIVHAGRSIGRFLTG 1079 AP00481 Ref, Kaliocin-1FFSASCVPGADKGQFPNLCRLCAGTG 1080 (synthetic) ENKCA AP00482 Ref, Thioninmutation KSCCRNTWARNCYNVCRLPGTISREIC 1081 (synthetic)AKKCRCKIISGTTCPSDYPK AP00484 Ref, Stomoxyn (insect,RGFRKHFNKLVKKVKHTISETAHVAK 1082 fly) DTAVIAGSGAAVVAAT AP00486 Ref,Cupiennin 1b GFGSLFKFLAKKVAKTVAKQAAKQG 1083 (spider) AKYIANKQME AP00487Ref, Cupiennin 1c GFGSLFKFLAKKVAKTVAKQAAKQG 1084 (spider) AKYIANKQTEAP00488 Ref, Cupiennin 1D GFGSLFKFLAKKVAKTVAKQAAKQG 1085 (spider)AKYVANKHME AP00489 Ref, Hipposin (fish) SGRGKTGGKARAKAKTRSSRAGLQFP 1086VGRVHRLLRKGNYAHRVGAGAPVYL AP00923 Ref, CarnobacteriocinAISYGNGVYCNKEKCWVNKAENKQA 1087 B1 (XXO, class IIa ITGIVIGGWASSLAGMGHbacteriocin, bacteria) AP00496 Ref, HP 2-20 (synthetic)AKKVFKRLEKLFSKIQNDK 1088 AP00497 Ref, Maximin H5 (toad)ILGPVLGLVSDTLDDVLGIL 1089 AP00498 Ref, rCRAMP (ratGLVRKGGEKFGEKLRKIGQKIKEFFQ 1090 cathelicidin) KLALEIEQ AP00500 Ref,S9-P18 (synthetic) KWKLFKKISKFLHLAKKF 1091 AP00501 Ref, L9-P18(synthetic) KWKLFKKILKFLHLAKKF 1092 AP00502 Ref, Clavaspirin (seaFLRFIGSVIHGIGHLVHHIGVAL 1093 squirt, tunicate) AP00503 Ref, human P-113DAKRHHGYKRKFH 1094 AP00504 Ref, human MUC7 20- LAHQKPFIRKSYKCLHKRCR 1095Mer AP00507 Ref, Nigrocin 2 (frog) GLLSKVLGVGKKVLCGVSGLC 1096 AP00508Ref, Nigrocin 1 (frog) GLLDSIKGMAISAGKGALQNLLKVAS 1097 CKLDKTC AP00509Ref, human Calcitermin VAIALKAAHYHTHKE 1098 AP00510 Ref, Dicynthaurin(sea ILQKAVLDCLKAAGSSLSKAAITAIYN 1099 peach) KIT AP00511 Ref, KIGAKIKIGAKIKIGAKIKIGAKI 1100 (synthetic) AP00516 Ref, Lycotoxin IIWLTALKFLGKHAAKHLAKQQLSKL 1101 (spider) AP00517 Ref, Lycotoxin IIKIKWFKTMKSIAKFIAKEQMKKHLGGE 1102 (spider) AP00518 Ref, Ib-AMP3 (plantQYRHRCCAWGPGRKYCKRWC 1103 defensin, balsam) AP00519 Ref, Ib-AMP4 (plantEWGRRCCGWGPGRRYCRRWC 1104 defensin, balsam) AP00521 Ref, Dhvar4(synthetic) KRLFKKLLFSLRKY 1105 AP00522 Ref, Dhvar5 (synthetic)LLLFLLKKRKKRKY 1106 AP00525 Ref, Maximin H2 (toad) ILGPVLSMVGSALGGLIKKI1107 AP00526 Ref, Maximin H3 (toad) ILGPVLGLVGNALGGLIKKI 1108 AP00527Ref, Maximin H4 (toad) ILGPVISKIGGVLGGLLKNL 1109 AP00528 Ref, Anionicpeptide DDDDDD 1110 SAAP (sheep) AP00530 Ref, Lantibiotic EricinVLSKSLCTPGCITGPLQTCYLCFPTFA 1111 A (bacteria) KC AP00531 Ref, KenojeininI (sea GKQYFPKVGGRLSGKAPLAAKTHRRL 1112 skate) KP AP00532 Ref, Lunatusin(plant, KTCENLADTFRGPCFATSNC 1113 ZZHp) AP00533 Ref, Fallaxin (frog)GVVDILKGAAKDIAGHLASKVMNKL 1114 AP00534 Ref, Tu-AMP 2KSCCRNTTARNCYNVCRIPG 1115 (TuAMP2, thionin-like antimicrobial peptides,plant defensin, tulip) AP00535 Ref, Pilosulin 1 (Myr bGLGSVFGRLARILGRVIPKVAKKLGPK 1116 I) (Australian ants)VAKVLPKVMKEAIPMAVEMAKSQEE QQPQ AP00536 Ref, Luxuriosin (insect)SVRTQDNAVNRQIFGSNGPYRDFQLS 1117 DCYLPLETNPYCNEWQFAYHWNNALMDCERAIYHGCNRTRNNFITLTACKN QAGPICNRRRH AP00537 Ref, SAMP H1 (fish,AEVAPAPAAAAPAKAPKKKAAAKPK 1118 Atlantic salmon) KAGPS AP00538 Ref,Halocidin (dimer WLNALLHHGLNCAKGVLA 1119 Hal18 + Hal15) (tunicate)AP00539 Ref, AOD (American GFGCPWNRYQCHSHCRSIGRLGGYCA 1120 oysterdefensin, animal GSLRLTCTCYRS defensin) AP00540 Ref, PentadactylinGLLDTLKGAAKNVVGSLASKVMEKL 1121 (frog) AP00541 Ref, Polybia-MPIIDWKKLLDAAKQIL 1122 (insect, social wasp) AP00542 Ref, Polybia-CP(insect, ILGTILGLLKSL 1123 social wasp) AP00543 Ref, Ocellatin-1 (XXA,GVVDILKGAGKDLLAHLVGKISEKV 1124 frog) AP00544 Ref, Ocellatin-2 (XXA,GVLDIFKDAAKQILAHAAEKQI 1125 frog) AP00545 Ref, Ocellatin-3 (frog)GVLDILKNAAKNILAHAAEQI 1126 AP00548 Ref, CMAP 27 (chickenRFGRFLRKIRRFRPKVTITIQGSARFG 1127 myeloid antimicrobial peptide 27, birdcathelicidin, chicken cathelicidin) AP00550 Ref, Tu-AMP-1KSCCRNTVARNCYNVCRIPGTPRPVC 1128 (TuAMP 1, thionin-likeAATCDCKLITGTKCPPGYEK antimicrobial peptides, plant defensin, tulip)AP00551 Ref, Combi-2 FRWWHR 1129 (synthetic) AP00552 Ref, Maximin 9(frog) GIGRKFLGGVKTTFRCGVKDFASKHLY 1130 AP00554 Ref, S1 moricin (insect)GKIPVKAIKKAGAAIGKGLRAINIAST 1131 AHDVYSFFKPKHKKK AP00555 Ref, Parasin I(catfish) KGRGKQGGKVRAKAKTRSS 1132 AP00556 Ref, Kassinatuerin-1GFMKYIGPLIPHAVKAISDLI 1133 (frog) AP00557 Ref, Fowlicidin-1RVKRVWPLVIRTVIAGYNLYRAIKKK 1134 (chCATH-1, bird cathelicidin, chickencathelicidin) AP00559 Ref, Eryngin ATRVVYCNRRSGSVVGGDDTVYYEG 1135(mushroom, fungi) AP00560 Ref, Dendrocin (plant,TTLTLHNLCPYPVWWLVTPNNGGFPII 1136 bamboo) DNTPVVLG AP00561 Ref, Coconutantifungal EQCREEEDDR 1137 peptide (plant) AP00562 Ref, Pandinin 1(African GKVWDWIKSAAKKIWSSEPVSQLKG 1138 scorpion) QVLNAAKNYVAEKIGATPTAP00563 Ref, White cloud bean KTCENLADTFRGPCFATSNCDDHCKN 1139 defensin(plant KEHLLSGRCRDDFRCWCTRNC defensin) AP00564 Ref, Dybowskin-1FLIGMTHGLICLISRKC 1140 (frog) AP00565 Ref, Dybowskin-2 FLIGMTQGLICLITRKC1141 (frog) AP00566 Ref, Dybowskin-3 GLFDVVKGVLKGVGKNVAGSLLEQL 1142(frog) KCKLSGGC AP00567 Ref, Dybowskin-4 VWPLGLVICKALKIC 1143 (frog)AP00568 Ref, Dybowskin-5 GLFSVVTGVLKAVGKNVAKNVGGSL 1144 (frog)LEQLKCKKISGGC AP00569 Ref, Dybowskin-6 FLPLLLAGLPLKLCFLFKKC 1145 (frog)AP00570 Ref, Pleurain-A1 (frog) SIITMTKEAKLPQLWKQIACRLYNTC 1146 AP00571Ref, Pleurain-A2 (frog) SIITMTKEAKLPQSWKQIACRLYNTC 1147 AP00574 Ref,Esculentin-IGRa GLFSKFAGKGIKNLIFKGVKHIGKEVG 1148 (frog)MDVIRTGIDVAGCKIKGEC AP00575 Ref, Brevinin-2GRaGLLDTFKNLALNAAKSAGVSVLNSLS 1149 (frog) CKLSKTC AP00576 Ref,Brevinin-2GRb GVLGTVKNLLIGAGKSAAQSVLKTLS 1150 (frog) CKLSNDC AP00577Ref, Brevinin-2GRc GLFTLIKGAAKLIGKTVAKEAGKTGLE 1151 (frog) LMACKITNQCAP00578 Ref, Brevinin-1GRa FLPLLAGLAANFLPKIFCKITKKC 1152 (frog) AP00579Ref, Nigrocin-2GRa GLLSGILGAGKHIVCGLSGLC 1153 (frog) AP00580 Ref,Nigrocin-2GRb GLFGKILGVGKKVLCGLSGMC 1154 (frog) AP00581 Ref,Nigrocin-2GRc GLLSGILGAGKNIVCGLSGLC 1155 (frog) AP00582 Ref,Brevinin-2GHa GFSSLFKAGAKYLLKSVGKAGAQQLA 1156 (frog) CKAANNCA AP00583Ref, Brevinin-2GHb GVITDALKGAAKTVAAELLRKAHCKL 1157 (frog) TNSC AP00584Ref, Guentherin (frog) VIDDLKKVAKKVRRELLCKKHHKKLN 1158 AP00585 Ref,Brevinin-2GHc SIWEGIKNAGKGFLVSILDKVRCKVA 1159 (frog) GGCNP AP00586 Ref,Temporin-GH FLPLLFGAISHLL 1160 (frog) AP00587 Ref, Brevinin-2TSaGIMSLFKGVLKTAGKHVAGSLVDQLK 1161 (frog) CKITGGC AP00588 Ref,Brevinin-1TSa FLGSIVGALASALPSLISKIRN 1162 (frog) AP00589 Ref,Temporin-1TSa FLGALAKIISGIF 1163 (frog) AP00593 Ref, Brevinin-1CSaFLPILAGLAAKIVPKLFCLATKKC 1164 (frog) AP00594 Ref, Temporin-1CSaFLPIVGKLLSGLL 1165 (frog) AP00595 Ref, Temporin-1CSb FLPIIGKLLSGLL 1166(frog) AP00596 Ref, Temporin-1CSc FLPLVTGLLSGLL 1167 (frog) AP00597 Ref,Temporin-1CSd NFLGTLVNLAKKIL 1168 (frog) AP00598 Ref, Temporin-1SPbFLSAITSLLGKLL 1169 (frog) AP00599 Ref, Brevinin-2-relatedGIWDTIKSMGKVFAGKILQNL 1170 (frog) AP00600 Ref, Odorranain-HPGLLRASSVWGRKYYVDLAGCAKA 1171 (frog) AP00601 Ref, Brevinin-1DYaFLSLALAALPKFLCLVFKKC 1172 (frog) AP00602 Ref, Brevinin-1DYbFLSLALAALPKLFCLIFKKC 1173 (frog) AP00603 Ref, Brevinin-1DYcFLPLLLAGLPKLLCLFFKKC 1174 (frog) AP00607 Ref, Brevinin-2DYbGLFDVVKGVLKGAGKNVAGSLLEQL 1175 (frog) KCKLSGGC AP00608 Ref,Brevinin-2DYc GLFDVVKGVLKGVGKNVAGSLLEQL 1176 (frog) KCKLSGGC AP00609Ref, Brevinin-2DYd GIFDVVKGVLKGVGKNVAGSLLEQLK 1177 (frog) CKLSGGCAP00610 Ref, Brevinin-2DYe GLFSVVTGVLKAVGKNVAKNVGGSL 1178 (frog)LEQLKCKISGGC AP00611 Ref, Temporin-1DYa FIGPIISALASLFG 1179 (frog)AP00615 Ref, Palustrin-1b (frog) ALFSILRGLKKLGNMGQAFVNCKIYK 1180 KCAP00616 Ref, Palustrin-1c (frog) ALSILRGLEKLAKMGIALTNCKATKKC 1181AP00617 Ref, Palustrin-1d (frog) ALSILKGLEKLAKMGIALTNCKATKKC 1182AP00619 Ref, Palustrin-2b (frog) GFFSTVKNLATNVAGTVIDTLKCKVT 1183 GGCRSAP00620 Ref, Palustrin-2c (frog) GFLSTVKNLATNVAGTVIDTLKCKVT 1184 GGCRSAP00621 Ref, Palustrin-3a (frog) GIFPKIIGKGIKTGIVNGIKSLVKGVGM 1185KVFKAGLNNIGNTGCNEDEC AP00622 Ref, Palustrin-3b (frog)GIFPKIIGKGIKTGIVNGIKSLVKGVGM 1186 KVFKAGLSNIGNTGCNEDEC AP00624 Ref,human ALL-38 (an ALLGDFFRKSKEKIGKEFKRIVQRIKD 1187 LL-37 analog releasedFLRNLVPRTES from its precursor hCAP-18 by gastricsin in vivo) AP00625Ref, human KR-20 KRIVQRIKDFLRNLVPRTES 1188 (KR20 from LL-37) AP00626Ref, human KS-30 KSKEKIGKEFKRIVQRIKDFLRNLVPR 1189 (KS30 from LL-37) TESAP00627 Ref, human RK-31 RKSKEKIGKEFKRIVQRIKDFLRNLVP 1190 (RK31 fromLL-37) RTES AP00628 Ref, human LL-23 LLGDFFRKSKEKIGKEFKRIVQR 1191 (LL23from LL-37) AP00629 Ref, human LL-29 LLGDFFRKSKEKIGKEFKRIVQRIKDFLR 1192(LL29 from LL-37) AP00630 Ref, Amoeba peptide GEILCNLCTGLINTLENLLTTKGAD1193 (protozoan para AP00631 Ref, Mundticin KYYGNGVSCNKKGCSVDWGKAIGIIG1194 (bacteria) NNSAANLATGGAAGWSK AP00638 Ref, Citropin 2.1 (frog)GLIGSIGKALGGLLVDVLKPKL 1195 AP00639 Ref, Citropin 2.1.3GLIGSIGKALGGLLVDVLKPKLQAAS 1196 (frog) AP00640 Ref, Maculatin 1.3GLLGLLGSVVSHVVPAIVGHF 1197 (frog) AP00641 Ref, Pardaxin 1GFFALIPKIISSPLFKTLLSAVGSALSSS 1198 (Pardaxin P-1, Pardaxin GEQE P1, Pa1,flat fish) AP00642 Ref, Pardaxin 2 GFFALIPKIISSPIFKTLLSAVGSALSSS 1199(Pardaxin P-2, Pardaxin GGQE P2, Pa2, flat fish) AP00643 Ref, Pardaxin 3GFFAFIPKIISSPLFKTLLSAVGSALSSS 1200 (Pardaxin P-3, Pardaxin GEQE P3, Pa3,flat fish) AP00645 Ref, Pardaxin 5 GFFAFIPKIISSPLFKTLLSAVGSALSSS 1201(Pardaxin P-5, Pardaxin GDQE P5, Pa5, flat fish) AP00647 Ref,Brevinin-1PLb FLPLIAGLAANFLPKIFCAITKKC 1202 (frog) AP00648 Ref,Brevinin-1PLc FLPVIAGVAAKFLPKIFCAITKKC 1203 (frog) AP00649 Ref,Esculentin-1PLa GLFPKINKKKAKTGVFNIIKTVGKEAG 1204 (frog)MDLIRTGIDTIGCKIKGEC AP00650 Ref, Esculentin-1PLbGIFTKINKKKAKTGVFNIIKTIGKEAG 1205 (frog) MDVIRAGIDTISCKIKGEC AP00651 Ref,Esculentin-2PLa GLFSILKGVGKIALKGLAKNMGKMGL 1206 (frog) DLVSCKISKECAP00652 Ref, Ranatuerin-2PLa GIMDTVKNVAKNLAGQLLDKLKCKIT 1207 (frog) ACAP00653 Ref, Ranatuerin-2PLb GIMDTVKNAAKDLAGQLLDKLKCRIT 1208 (frog) GCAP00654 Ref, Ranatuerin-2PLc GLLDTIKNTAKNLAVGLLDKIKCKMT 1209 (frog) GCAP00655 Ref, Ranatuerin-2PLd GIMDSVKNVAKNIAGQLLDKLKCKIT 1210 (frog) GCAP00656 Ref, Ranatuerin-2PLe GIMDSVKNAAKNLAGQLLDTIKCKIT 1211 (frog) ACAP00657 Ref, Ranatuerin-2PLf GIMDTVKNAAKDLAGQLDKLKCRITGC 1212 (frog)AP00658 Ref, Temporin-1PLa FLPLVGKILSGLI 1213 (frog) AP00659 Ref,Ranatuerin 5 (frog) FLPIASLLGKYL 1214 AP00661 Ref, Esculentin-2LGILSLFTGGIKALGKTLFKMAGKAGA 1215 (frog) EHLACKATNQC AP00662 Ref,Esculentin-2B GLFSILRGAAKFASKGLGKDLTKLGV 1216 (ESC2B-RANBE, frog)DLVACKISKQC AP00663 Ref, Esculentin-2P GFSSIFRGVAKFASKGLGKDLARLGV 1217(frog) NLVACKISKQC AP00664 Ref, Peptide A1 (frog) FLPAIAGILSQLF 1218AP00665 Ref, Peptide B9 (frog) FLPLIAGLIGKLF 1219 AP00666 Ref, PG-L(frog) EGGGPQWAVGHFM 1220 AP00667 Ref, PG-KI (frog) EPHPDEFVGLM 1221AP00668 Ref, PG-KII (frog) EPNPDEFVGLM 1222 AP00669 Ref, PG-KIII (frog)EPHPNEFVGLM 1223 AP00670 Ref, PG-SPI (frog) EPNPDEFFGLM 1224 AP00660Ref, Pandinin 2 (African FWGALAKGALKLIPSLFSSFSKKD 1225 scorpion) AP00671Ref, PG-SPII (frog) EPNPNEFFGLM 1226 AP00673 Ref, Lantibiotic Ericin SWKSESVCTPGCVTGVLQTCFLQTITC 1227 (bacteria NCHISK AP00674 Ref,Lantibiotic Ericin VLSKSLCTPGCITGPLQTCYLCFPTFA 1228 A (bacteria KCAP00675 Ref, Human beta FELDRICGYGTARCRKKCRSQEYRIGR 1229 defensin 4(HBD-4, CPNTYACCLRKWDESLLNRTKP HBD4, human defensin) AP00676 Ref, RL-37(RL37, RLGNFFRKVKEKIGGGLKKVGQKIKD 1230 monkey cathelicidin) FLGNLVPRTASAP00677 Ref, CAP11 (Guinea pig GLRKKFRKTRKRIQKLGRKIGKTGRK 1231cathelicidin) VWKAWREYGQIPYPCRI AP00678 Ref, Canine cathelicidinRLKELITTGGQKIGEKIRRIGQRIKDFF 1232 (K9CATH) (dog) KNLQPREEKS AP00679 Ref,Esculentin 2VEb GLFSILKGVGKIAIKGLGKNLGKMGL 1233 (frog) DLVSCKISKECAP00680 Ref, SMAP-34 (sheep GLFGRLRDSLQRGGQKILEKAERIWC 1234cathelicidin) KIKDIFR AP00681 Ref, OaBac5 (sheepRFRPPIRRPPIRPPFRPPFRPPVRPPIRPP 1235 cathelicidin) FRPPFRPPIGPFP AP00682Ref, OaBac6 (sheep RRLRPRHQHFPSERPWPKPLPLPLPRP 1236 cathelicidin)GPRPWPKPLPLPLPRPGLRPWPKPL AP00683 Ref, OaBac7.5 (sheepRRLRPRRPRLPRPRPRPRPRPRSLPLPR 1237 cathelicidin)PQPRRIPRPILLPWRPPRPIPRPQIQPIPR WL AP00684 Ref, OaBac11 (sheepRRLRPRRPRLPRPRPRPRPRPRSLPLPR 1238 cathelicidin)PKPRPIPRPLPLPRPRPKPIPRPLPLPRP RPRRIPRPLPLPRPRPRPIPRPLPLPQPQ PSPIPRPLAP00685 Ref, Ranatuerin 2VEb GIMDTVKGVAKTVAASLLDKLKCKIT 1239 (frog) GCAP00686 Ref, eCATH-1 (horse KRFGRLAKSFLRMRILLPRRKILLAS 1240cathelicidin) AP00687 Ref, eCATH-2 (horse KRRHWFPLSFQEFLEQLRRFRDQLPFP1241 cathelicidin) AP00688 Ref, eCATH-3 (horseKRFHSVGSLIQRHQQMIRDKSEATRH 1242 cathelicidin) GIRIITRPKLLLAS AP00689Ref, Prophenin-1 (pig AFPPPNVPGPRFPPPNFPGPRFPPPNFP 1243 cathelicidin)GPRFPPPNFPGPRFPPPNFPGPPFPPPIFP GPWFPPPPPFRPPPFGPPRFP AP00690 Ref,Prophenin-2 (pig AFPPPNVPGPRFPPPNVPGPRFPPPNFP 1244 cathelicidin)GPRFPPPNFPGPRFPPPNFPGPPFPPPIFP GPWFPPPPPFRPPPFGPPRFP AP00691 Ref,HFIAP-1 (hagfish GFFKKAWRKVKHAGRRVLDTAKGV 1245 cathelicidin)GRHYVNNWLNRYR AP00692 Ref, HFIAP-3 (hagfish GWFKKAWRKVKNAGRRVLKGVGIH1246 cathelicidin) YGVGLI AP00693 Ref, Trout cath (fishRICSRDKNCVSRPGVGSIIGRPGGGSLI 1247 cathelicidin)GRPGGGSVIGRPGGGSPPGGGSFNDEF IRDHSDGNRFA AP00694 Ref, MRP (melittin-AIGSILGALAKGLPTLISWIKNR 1248 related peptide) AP00695 Ref, Temporin-1TGaFLPILGKLLSGIL 1249 (frog) AP00696 Ref, Dahlein 1.1 (frog) GLFDIIKNIVSTL1250 AP00697 Ref, Dahlein 1.2 (frog) GLFDIIKNIFSGL 1251 AP00698 Ref,Dahlein 4.1 (frog) GLWQLIKDKIKDAATGFVTGIQS 1252 AP00699 Ref, Dahlein 4.2(frog) GLWQFIKDKLKDAATGLVTGIQS 1253 AP00700 Ref, Dahlein 4.3 (frog)GLWQFIKDKFKDAATGLVTGIQS 1254 AP00701 Ref, Dahlein 5.1 (frog)GLLGSIGNAIGAFIANKLKP 1255 AP00702 Ref, Dahlein 5.2 (frog)GLLGSIGNAIGAFIANKLKPK 1256 AP00703 Ref, Dahlein 5.3 (frog)GLLASLGKVLGGYLAEKLKP 1257 AP00704 Ref, Dahlein 5.4 (frog)GLLGSIGKVLGGYLAEKLKPK 1258 AP00705 Ref, Dahlein 5.5 (frog)GLLASLGKVLGGYLAEKLKPK 1259 AP00706 Ref, Dahlein 5.6 (frog)GLLASLGKVFGGYLAEKLKPK 1260 AP00709 Ref, Mytilus defensinGFGCPNDYPCHRHCKSIPGRAGGYCG 1261 (mytilin) A (mollusc) GAHRLRCTCYRAP00711 Ref, Mussel defensin GFGCPNNYACHQHCKSIRGYCGGYC 1262 MGD2AGWFRLRCTCYRCG AP00712 Ref, scorpion defensin GFGCPLNQGACHRHCRSIRRRGGYCA1263 GFFKQTCCYRN AP00713 Ref, Androctonus GFGCPFNQGACHRHCRSIRRRGGYCA1264 defensin GLFKQTCTCYR AP00714 Ref, OrinthodorosGYGCPFNQYQCHSHCSGIRGYKGGYC 1265 defensin A (soft ticks) KGTFKQTCKCYAP00715 Ref, VaD1 (plant RTCMKKEGWGKCLIDTTCAHSCKNR 1266 defensin)GYIGGNCKGMTRTCYCLVNC AP00722 Ref, Cryptonin (insect,GLLNGLALRLGKRALKKIIKRLCR 1267 cicada) AP00723 Ref, Decoralin (insect)SLLSLLRKLIT 1268 AP00724 Ref, RTD-2 (rhesus RCLCRRGVCRCLCRRGVC 1269theta-defensin-2, minidefensin, XXC, BBS, lectin, ZZHa) AP00725 Ref,RTD-3 (rhesus RCICTRGFCRCICTRGFC 1270 theta-defensin-3, minidefensin,XXC, BBS, lectin, ZZHa) AP00726 Ref, Combi-1 RRWWRF 1271 (synthetic)AP00748 Ref, Gm pro-rich pept1 DIQIPGIKKPTHRDIIIPNWNPNVRTQP 1272(insect) WQRFGGNKS AP00749 Ref, Gm anionic pept 1EADEPLWLYKGDNIERAPTTADHPILP 1273 (insect) SIIDDVKLDPNRRYA AP00750 Ref,Gm pro-rich pept 2 EIRLPEPFRFPSPTVPKPIDIDPILPHPWS 1274 (insect)PRQTYPIIARRS AP00752 Ref, Gm defensin-like DKLIGSCVWGATNYTSDCNAECKRR1275 peptide (insect) GYKGGHCGSFWNVNCWCEE AP00753 Ref, GmVQETQKLAKTVGANLEETNKKLAPQI 1276 apolipophoricin (insect)KSAYDDFVKQAQEVQKKLHEAASKQ AP00754 Ref, Gm anionic pept2ETESTPDYLKNIQQQLEEYTKNFNTQV 1277 (insect) QNAFDSDKIKSEVNNFIESLGKILNTEKKEAPK AP00755 Ref, Gm cecropin D- ENFFKEIERAGQRIRDAIISAAPAVETL 1278like pept, insect AQAQKIIKGGD AP00756 Ref, Dermaseptin-B6ALWKDILKNAGKAALNEINQLVNQ 1279 (DRS-B6, DRS B6, XXA, frog) AP00759 Ref,Phylloseptin-O1 FLSLIPHAINAVSTLVHHSG 1280 (PLS-O1, Phylloseptin- 4,PS-4, XXA, frog) AP00760 Ref, Phylloseptin-O2 FLSLIPHAINAVSAIAKHS 1281(PLS-O2, Phylloseptin- 5, PS-5, XXA, frog) AP00761 Ref, Phylloseptin-6SLIPHAINAVSAIAKHF 1282 (Phylloseptin-H4, PLS- H4, PS-6, XXA, frog)AP00762 Ref, Phylloseptin-7 FLSLIPHAINAVSAIAKHF 1283 (Phylloseptin-H5,PLS- H5, PS-7, XXA, frog) AP00763 Ref, Dermaseptin DPh-1GLWSTIKNVGKEAAIAAGKAALGAL 1284 (XXA, frog) AP00764 Ref, Dermaseptin-S9GLRSKIWLWVLLMIWQESNKFKKM 1285 (DRS-S9, DRS S9, frog) AP00765 Ref, Humansalvic MHDFWVLWVLLEYIYNSACSVLSATS 1286 SVSSRVLNRSLQVKVVKITN AP00766 Ref,Gassericin A IYWIADQFGIHLATGTARKLLDAMAS 1287 (XXC, XXD2, class IVGASLGTAFAAILGVTLPAWALAAAGA bacteriocin, Gram- LGATAA positive bacteria)AP00767 Ref, Circularin A (XXC, VAGALGVQTAAATTIVNVILNAGTLV 1288 class IVbacteriocin, TVLGIIASIASGGAGTLMTIGWATFKA Gram-positive bacteria)TVQKLAKQSMARAIAY AP00768 Ref, Closticin 574 PNWTKIGKCAGSIAWAIGSGLFGGAK1289 (bacteria) LIKIKKYIAELGGLQKAAKLLVGATT WEEKLHAGGYALINLAAELTGVAGIQANCF AP00769 Ref, Caerin 1.11 (XXA, GLLGAMFKVASKVLPHVVPAITEHF 1290 frog)AP00770 Ref, Maculatin 1.4 GLLGLLGSVVSHVLPAITQHL 1291 (XXA, frog)AP00771 Ref, Magainin 1 (frog) GIGKFLHSAGKFGKAFVGEIMKS 1292 AP00772 Ref,Oxyopinin 1 FRGLAKLLKIGLKSFARVLKKVLPKA 1293 (spider)AKAGKALAKSMADENAIRQQNQ AP00773 Ref, Oxyopinin 2aGKFSVFGKILRSIAKVFKGVGKVRKQF 1294 (spider) KTASDLDKNQ AP00774 Ref,Oxyopinin 2b GKFSGFAKILKSIAKFFKGVGKVRKGF 1295 (spider) KEASDLDKNQAP00775 Ref, Oxyopinin 2c GKLSGISKVLRAIAKFFKGVGKARKQ 1296 (spider)FKEASDLDKNQ AP00776 Ref, Oxyopinin 2d GKFSVFSKILRSIAKVFKGVGKVRKGF 1297(spider) KTASDLDKNQ AP00777 Ref, NRC-1 (XXA, fish,GKGRWLERIGKAGGIIIGGALDHL 1298 gene predicted) AP00778 Ref, NRC-2 (XXA,fish, WLRRIGKGVKIIGGAALDHL 1299 gene predicted) AP00779 Ref, NRC-3 (XXA,fish, GRRKRKWLRRIGKGVKIIGGAALDHL 1300 gene predicted) AP00781 Ref, NRC-5(XXA, fish, FLGALIKGAIHGGRFIHGMIQNHH 1301 gene predicted) AP00782 Ref,NRC-6 (XXA, fish, GWGSIFKHGRHAAKHIGHAAVNHYL 1302 gene predicted) AP00783Ref, NRC-7 (XXA, fish, RWGKWFKKATHVGKHVGKAALTAYL 1303 gene predicted)AP00784 Ref, NRC-10 (XXA, FFRLLFHGVHHVGKIKPRA 1304 fish, gene predicted)AP00785 Ref, NRC-11 (XXA, GWKSVFRKAKKVGKTVGGLALDHYL 1305 fish, genepredicted) AP00786 Ref, NRC-12 (XXA, GWKKWFNRAKKVGKTVGGLAVDHYL 1306fish, gene predicted) AP00787 Ref, NRC-13 (XXA, GWRLLLKKAEVKTVGKLALKHYL1307 fish, gene predicted) AP00788 Ref, NRC-14 (XXA,AGWGSIFKHIFKAGKFIHGAIQAHND 1308 fish, gene predicted) AP00789 Ref,NRC-15 (XXA, GFWGKLFKLGLHGIGLLHLHL 1309 fish, gene predicted) AP00790Ref, NRC-16 (XXA, GWKKWLRKGAKHLGQAAIK 1310 fish, gene predicted) AP00791Ref, NRC-17 (XXA, GWKKWLRKGAKHLGQAAIKGLAS 1311 fish, gene predicted)AP00792 Ref, NRC-19 (XXA, FLGLLFHGVHHVGKWIHGLIHGHH 1312 fish, genepredicted) AP00793 Ref, Bombinin H2 IIGPVLGLVGSALGGLLKKI 1313 (XXA,frog) AP00794 Ref, Bombinin H3 (frog, IIGPVLGMVGSALGGLLKKI 1314 XXD,XXA) AP00795 Ref, Bombinin H7 (frog, ILGPILGLVSNALGGLL 1315 XXD, XXA)AP00796 Ref, Bombinin GH-1L IIGPVLGLVGKPLESLLE 1316 (XXA, toad) AP00797Ref, Bombinin GH-1D IIGPVLGLVGKPLESLLE 1317 (toad, XXD, XXA) AP00807Ref, Enterocin E-760 NRWYCNSAAGGVGGAAGCVLAGYV 1318 (bacteriocin,bacteria) GEAKENIAGEVRKGWGMAGGFTHNK ACKSFPGSGWASG AP00808 Ref, hepcidin(fish) CRFCCRCCPRMRGCGLCCRF 1319 AP00809 Ref, hepcidin TH1-5GIKCRFCCGCCTPGICGVCCRF 1320 (fish) AP00810 Ref, hepcidin TH2-3QSHLSLCRWCCNCCRSNKGC 1321 (fish) AP00811 Ref, human LEAP-2MTPFWRGVSLRPIGASCRDDSECITRL 1322 CRKRRCSLSVAQE AP00812 Ref, Enkelytin(cow) FAEPLPSEEEGESYSKEPPEMEKRYGG 1323 FM AP00732 Ref, Spheniscin-1SFGLCRLRRGSCAHGRCRFPSIPIGRCS 1324 (Sphe-1, avian defensin) RFVQCCRRVWAP00733 Ref, Organgutan LLGDFFRKAREKIGEEFKRIVQRIKDFL 1325 ppyLL-37(Great Ape, RNLVPRTES primate cathelicidin) AP00734 Ref, Gibbon hmdSL-37SLGNFFRKARKKIGEEFKRIVQRIKDF 1326 (hylobatidae, primate LQHLIPRTEAcathelicidin) AP00735 Ref, pobRL-37 RLGNFFRKAKKKIGRGLKKIGQKIKDF 1327(cercopithecidae, LGNLVPRTES primate cathelicidin) AP00736 Ref, cjaRL-37(primate RLGDILQKAREKIEGGLKKLVQKIKDF 1328 cathelicidin) FGKFAPRTESAP00737 Ref, Plasticin PBN2KF GLVTSLIKGAGKLLGGLFGSVTG 1329 (XXA,DRP-PBN2, frog) AP00738 Ref, Plasticin ANCKF GLVTGLLKTAGKLLGDLFGSLTG1330 (XXA, synthetic) AP00739 Ref, Plasticin PD36KFGVVTDLLKTAGKLLGNLFGSLSG 1331 (XXA, synthetic) AP00740 Ref, PlasticinPD36K GVVTDLLKTAGKLLGNLVGSLSG 1332 (XXA, synthetic) AP00741 Ref, ChickenPITYLDAILAAVRLLNQRISGPCILRLR 1333 cathelicidin-B1 (birdEAQPRPGWVGTLQRRREVSFLVEDGP cathelicidin) CPPGVDCRSCEPGALQHCVGTVSIEQQPTAELRCRPLRPQ AP00742 Ref, Chicken gallinacinMRILYLLLSVLFVVLQGVAGQPYFSSP 1334 4 (Gal 4) IHACRYQRGVCIPGPCRWPYYRVGSCGSGLKSCCVRNRWA AP00743 Ref, Chicken gallinacinMKILCFFIVLFVAVHGAVGFSRSPRYH 1335 7 (Gal 7) MQCGYRGTFCTPGKCPYGNAYLGLCRPKYSCCRWL AP00744 Ref, Chicken gallinacin MQILPLLFAVLLLMLRAEPGLSLARGL1336 9 (Gal 9) PQDCERRGGFCSHKSCPPGIGRIGLCS KEDFCCRSRWYS AP00745 Ref,Chicken LEAP-2 MTPFWRGVSLRPVGASCRDNSECITM 1337 (cLEAP-2) LCRKNRCFLRTASEAP00814 Ref, Caerulein GLGSILGKILNVAGKVGKTIGKVADA 1338 precursor-relatedVGNKE fragment Ea (CPRF-Ea, frog) AP00815 Ref, CaeruleinGLGSFLKNAIKIAGKVGSTIGKVADAI 1339 precursor-related GNKE fragment Eb(CPRF-Eb, frog) AP00816 Ref, Caerulein GLGSFFKNAIKIAGKVGSTIGKVADAI 1340precursor-related GNKE fragment Ec (CPRF-Ec, frog) AP00817 Ref,Temporin-1Oa FLPLLASLFSRLL 1341 (frog) AP00818 Ref, Temporin-1ObFLPLIGKILGTIL 1342 (frog) AP00819 Ref, Temporin-1Oc FLPLLASLFSRLF 1343(frog) AP00820 Ref, Temporin-1Od FLPLLASLFSGLF 1344 (frog) AP00821 Ref,Brevinin-20a (frog) GLFNVFKGLKTAGKHVAGSLLNQLK 1345 CKVSGGC AP00822 Ref,Brevinin-20b (frog) GIFNVFKGALKTAGKHVAGSLLNQLK 1346 CKVSGEC AP00824 Ref,Temporin-1Gb SILPTIVSFLSKFL 1347 (XXA, frog) AP00825 Ref, Temporin-1GcSILPTIVSFLTKFL 1348 (XXA, frog) AP00826 Ref, Temporin-1Gd FILPLIASFLSKFL1349 (XXA, frog) AP00827 Ref, Ranatuerin-1Ga SMISVLKNLGKVGLGFVACKVNKQC1350 (frog) AP00829 Ref, Ranalexin-1G FLGGLMKIIPAAFCAVTKKC 1351 (frog)AP00830 Ref, Ranatuerin-2G GLLLDTLKGAAKDIAGIALEKLKCKIT 1352 (frog) GCKPAP00831 Ref, Odorranain-NR GLLSGILGAGKHIVCGLTGCAKA 1353 (frog) AP00832Ref, Maximin H1 ILGPVISTIGGVLGGLLKNL 1354 (XXA, toad) AP00834 Ref, G.mellonella KVNANAIKKGGKAIGKGFKVISAAST 1355 moricin-like peptide AAHDVYEHIKNRRH (Gm-mlpA, insect) AP00835 Ref, G. mellonellaGKIPVKAIKKGGQIIGKALRGINIASTA 1356 moricin-like peptide B HDIISQFKPKKKKNH(Gm-mlpB, insect) AP00836 Ref, G. mellonella KVPIGAIKKGGKIIKKGLGVIGAAGTA1357 moricin-like peptide C1 HEVYSHVKNRH (Gm-mlpC1, insect) AP00837 Ref,G. mellonella KVPIGAIKKGGKIIKKGLGVLGAAGTA 1358 moricin-like peptide C2HEVYNHVRNRQ (Gm-mlpC2, insect) AP00838 Ref, G. mellonellaKVPIGAIKKGGKIIKKGLGVIGAAGTA 1359 moricin-like peptide C3 HEVYSHVKNRQ(Gm-mlpC3, insect) AP00839 Ref, G. mellonella KVPVGAIKKGGKAIKTGLGVVGAAGT1360 moricin-like peptide AHEVYSHIRNRH C4/C5 (Gm-mlpC4/C5, insect)AP00840 Ref, G. mellonella KGIGSALKKGGKIIKGGLGALGAIGTG 1361 moricin-likepeptide D QQVYEHVQNRQ (Gm-mlpD, insect) AP00841 Ref, Enterocin A (EntA,TTHSGKYYGNGVYCTKNKCTVDWAK 1362 class IIA bacteriocin,ATTCIAGMSIGGFLGGAIPGKC i.e. pediocin-like peptide, bacteria) AP00842Ref, Divercin V41 TKYYGNGVYCNSKKCWVDWGQASG 1363 (DvnV41, class IIaCIGQTVVGGWLGGAIPGKC bacteriocin, pediocin- like peptide, bacteria.DvnRV41 is the recombinant form) AP00843 Ref, Divergicin M35TKYYGNGVYCNSKKCWVDWGTAQG 1364 (class IIa bacteriocin,CIDVVIGQLGGGIPGKGKC pediocin-like peptide, bacteria) AP00844 Ref,Coagulin KYYGNGVTCGKHSCSVDWGKATTCII 1365 (bacteriocin, pediocin-NNGAMAWATGGHQGTHKC like peptide, bacteria) AP00845 Ref, Listeriocin 743AKSYGNGVHCNKKKCWVDWGSAISTI 1366 (class IIa bacteriocin,GNNSAANWATGGAAGWKS pediocin-like peptide, bacteria) AP00846 Ref,Mundticin KS KYYGNGVSCNKKGCSVDWGKAIGIIG 1367 (enterocin CRL35,NNSAANLATGGAAGWKS mundticin ATO6, mundticin QU2, class IIa bacteriocin,pediocin-like peptide, bacteria) AP00847 Ref, Sakacin 5XKYYGNGLSCNKSGCSVDWSKAISIIGN 1368 (Sak5X, class IIa NAVANLTTGGAAGWKSbacteriocin, pediocin- like peptide, bacteria) AP00848 Ref, Leucocin C(class KNYGNGVHCTKKGCSVDWGYAWAN 1369 IIa bacteriocin,IANNSVMNGLTGGNAGWHN pediocin-like peptide, bacteria) AP00849 Ref,Lactococcin TSYGNGVHCNKSKCWIDVSELETYKA 1370 MMFII (class IIa GTVSNPKDILWbacteriocin, pediocin- like peptide, bacteria) AP00850 Ref, Sakacin G(SakG, KYYGNGVSCNSHGCSVNWGQAWTC 1371 class IIa bacteriocin,GVNHLANGGHGVC pediocin-like peptide, bacteria) AP00851 Ref, Plantaricin423 KYYGNGVTCGKHSCSVNWGQAFSCS 1372 (class IIa bacteriocin, VSHLANFGHGKCpediocin-like peptide, bacteria) AP00852 Ref, Plantaricin C19KYYGNGLSCSKKGCTVNWGQAFSCG 1373 (class IIa bacteriocin, VNRVATAGHHKCpediocin-like peptide, bacteria) AP00853 Ref, Enterocin P (EntP,ATRSYGNGVYCNNSKCWVNWGEAK 1374 class IIa bacteriocin,ENIAGIVISGWASGLAGMGH pediocin-like peptide, bacteria) AP00854 Ref,Bacteriocin 31 ATYYGNGLYCNKQKCWVDWNKASR 1375 (Bac 31, Bac31, classEIGKIIVNGWVQHGPWAPR IIa bacteriocin, pediocin-like peptide, bacteria)AP00855 Ref, MSI-78 (XXA, GIGKFLKKAKKFGKAFVKILKK 1376 synthetic) AP00856Ref, MSI-594 (XXA, GIGKFLKKAKKGIGAVLKVLTTGL 1377 synthetic) AP00857 Ref,Catestatin (human SSMKLSFRARAYGFRGPGPQL 1378 CHGA(352-372), human Cst)AP00858 Ref, Temporin D (XXA, LLPIVGNLLNSLL 1379 frog) AP00859 Ref,Temporin H (XXA, LSPNLLKSLL 1380 frog) AP00861 Ref, Brevinin-ALbFLPLAVSLAANFLPKLFCKITKKC 1381 (frog) AP00862 Ref, Brevinin 1E (frog)FLPLLAGLAANFLPKIFCKITKRC 1382 AP00863 Ref, Temporin-ALa FLPIVGKLLSGLSGLL1383 (XXA, frog) AP00864 Ref, Temporin 1ARa FLPIVGRLISGLL 1384 (XXA,frog) AP00865 Ref, Temporin 1AUa FLPIIGQLLSGLL 1385 (XXA, Temporin-1AUa) (frog) AP00866 Ref, Temporin 1Bya FLPIIAKVLSGLL 1386 (XXA,Temporin-1Bya, frog) AP00867 Ref, Temporin 1Ec FLPVIAGLLSKLF 1387 (XXA,frog) AP00869 Ref, Temporin 1Ja ILPLVGNLLNDLL 1388 (XXA, Temporin-1Ja,frog) AP00873 Ref, Temporin 1Pra ILPILGNLLNGLL 1389 (XXA, frog) AP00874Ref, Temporin 1VE FLPLVGKILSGLI 1390 (XXA, frog) AP00875 Ref, Temporin1Va FLSSIGKILGNLL 1391 (XXA, frog) AP00876 Ref, Temporin 1VbFLSIIAKVLGSLF 1392 (XXA, frog) AP00877 Ref, Brevinin-1Ja (frog)FLGSLIGAAIPAIKQLLGLKK 1393 AP00878 Ref, Brevinin-1BYaFLPILASLAAKFGPKLFCLVTKKC 1394 (frog) AP00884 Ref, Ixosin-B (tick)QLKVDLWGTRSGIQPEQHSSGKSDVR 1395 RWRSRY AP00885 Ref, Brevinin-1BYbFLPILASLAAKLGPKLFCLVTKKC 1396 (frog) AP00886 Ref, Brevinin-1BYcFLPILASLAATLGPKLLCLITKKC 1397 (frog) AP00887 Ref, Brevinin-2BYaGILSTFKGLAKGVAKDLAGNLLDKFK 1398 (frog) CKITGC AP00888 Ref, Brevinin-2BYbGIMDSVKGLAKNLAGKLLDSLKCKIT 1399 (frog) GC AP00891 Ref, Pilosulin 3 (Myrb IIGLVSKGTCVLVKTVCKKVLKQG 1400 III)(ants) AP00892 Ref, Pilosulin 4 (Myrb PDITKLNIKKLTKATCKVISKGASMCK 1401 IV)(ants) VLFDKKKQE AP00893 Ref,Pilosulin 5 (Myr b DVKGMKKAIKGILDCVIEKGYDKLAA 1402 III)(ants)KLKKVIQQLWE AP00894 Ref, Ocellatin 4 (XXA, GLLDFVTGVGKDIFAQLIKQI 1403frog) AP00895 Ref, OH-CATH (snake KRFKKFFKKLKNSVKKRAKKFFKKPR 1404cathelicidin, reptile VIGVSIPF cathelicidin, or elapid cathelicidins)AP00896 Ref, BF-CATH (snake KRFKKFFKKLKKSVKKRAKKFFKKPR 1405cathelicidin) VIGVSIPF AP00897 Ref, NA-CATH (snakeKRFKKFFKKLKNSVKKRAKKFFKKPK 1406 cathelicidin) VIGVTFPF AP00898 Ref,Temporin-1Sa FLSGIVGMLGKLF 1407 (XXA, frog) AP00899 Ref, Temporin-1SbFLPIVTNLLSGLL 1408 (XXA, frog) AP00900 Ref, Temporin-1Sc FLSHIAGFLSNLF1409 (XXA, frog) AP00913 Ref, Ib-AMP1 EWGRRCCGWGPGRRYCVRWC 1410 (IbAMP1,plant defensin) AP00914 Ref, Ib-AMP2 QYGRRCCNWGPGRRYCKRWC 1411 (IBAMP2,plant defensin) AP00915 Ref, Ee-CBP (EeCBP, QQCGRQAGNRRCANNLCCSQYGYCG1412 plant defensin, hevein- RTNEYCCTSQGCQSQCRRCG type, E. europaeuschitin-binding protein) AP00916 Ref, Pa-AMP1 AGCIKNGGRCNASAGPPYCCSSYCFQI1413 (PaAMP1, plant AGQSYGVCKNR defensin, C6 type) AP00917 Ref, Pa-AMP2ACIKNGGRCVASGGPPYCCSNYCLQI 1414 (PaAMP2, plant AGQSYGVCKKH defensin, C6type) AP00924 Ref, Ornithodoros GYGCPFNQYQCHSHCRGIRGYKGGY 1415 defensinB (soft ticks) CTGRFKQTCKCY AP00925 Ref, OrnithodorosGYGCPFNQYQCHSHCSGIRGYKGGYC 1416 defensin C (soft ticks) KGLFKQTCNCYAP00926 Ref, Ornithodoros GFGCPFNQYECHAHCSGVPGYKGGY 1417 defensin D(soft ticks) CKGLFKQTCNCY AP00927 Ref, ButyrivibriocinIYFIADKMGIQLAPAWYQDIVNWVSA 1418 AR10 (XXC, class IVGGTLTTGFAIIVGVTVPAWIAEAAAAF bacteriocin, gram- GIASA positive bacteria)AP00929 Ref, AS-48 (enterocin 4, ASLQFLPIAHMAKEFGIPAAVAGTVIN 1419 XXC,class IV VVEAGGWVTTIVSILTAVGSGGLSLLA bacteriocin or class IIdAAGRESIKAYLKKEIKKKGKRAVIAW bacteriocin, Gram- positive bacteria) AP00930Ref, Reutericin 6 (XXC, IYWIADQFGIHLATGTARKLLDAMAS 1420 XXD1, class IVGASLGTAFAAILGVTLPAWALAAAGA bacteriocin, Gram- LGATAA positive bacteria)AP00931 Ref, Uberolysin (XXC, LAGYTGIASGTAKKVVDAIDKGAAAF 1421 class IVbacteriocin, VIISIISTVISAGALGAVSASADFIILTVK Gram-positive bacteria)NYISRNLKAQAVIW AP00932 Ref, Acidocin B (XXC, IYWIADQFGIHLATGTARKLLDAVAS1422 class IV bacteriocin, GASLGTAFAAILGVTLPAWALAAAGA Gram-positivebacteria) LGATAA AP00980 Ref, Phormia defensin BATCDLLSGTGINHSACAAHCLLRGNR 1423 (insect defensin B) GGYCNRKGVCVCRNAP00990 Ref, Pth-St1 (plant RNCESLSHRFKGPCTRDSN 1424 defensin) AP00991Ref, Snakin-1 (StSN1, GSNFCDSKCKLRCSKAGLADRCLKYC 1425 plant defensin)GICCEECKCVPSGTYGNKHECPCYRD KKNSKGKSKCP AP00992 Ref, Snakin-2 (StSN2,YSYKKIDCGGACAARCRLSSRPRLCN 1426 plant defensin)RACGTCCARCNCVPPGTSGNTETCPC YASLTTHGNKRKCP AP00993 Ref, So-D2 (S.oleracea GIFSSRKCKTPSKTFKGICTRDSNCDTS 1427 defensin D2, plantCRYEGYPAGDCKGIRRRCMCSKPC defensin) AP00994 Ref, So-D6 (S. oleraceaGIFSNMYARTPAGYFRGP 1428 defensin D6, plant defensin) AP00997 Ref, NisinQ ITSISLCTPGCKTGVLMGCNLKTATCN 1429 (lantibiotic, CSVHVSK bacteriocins,bacteria) AP01008 Ref, Tachystatin A1 YSRCQLQGFNCVVRSYGLPTIPCCRGL 1430(BBS, horseshoe crabs) TCRSYFPGSTYGRCQRF AP01009 Ref, Tachystatin CDYDWSLRGPPKCATYGQKCRTWSPR 1431 (BBS, horseshoe crabs) NCCWNLRCKAFRCRPRAP01012 Ref, Latarcin 3a (Ltc3a, SWKSMAKKLKEYMEKLKQRA 1432 XXA, BBM,spider) AP01013 Ref, Latarcin 3b (Ltc3b, SWASMAKKLKEYMEKLKQRA 1433 XXA,BBM, spider) AP01014 Ref, Latarcin 4a (Ltc4a, GLKDKFKSMGEKLKQYIQTWKAKF1434 XXA, BBM, spider) AP01015 Ref, Latarcin 4b (Ltc4b,SLKDKVKSMGEKLKQYIQTWKAKF 1435 XXA, BBM, spider) AP01016 Ref, Latarcin 5(Ltc5, GFFGKMKEYFKKFGASFKRRFANLKK 1436 XXA, BBM, spider) RL AP01018 Ref,Latarcin 6a (Ltc6a, QAFQTFKPDWNKIRYDAMKMQTSLG 1437 BBM, spider) QMKKRFNLAP01019 Ref, Latarcin 7 (Ltc7, GETFDKLKEKLKTFYQKLVEKAEDLK 1438 BBM,spider) GDLKAKLS AP01049 Ref, Kalata B2 (plantVCGETCFGGTCNTPGCSCTWPICTRD 1439 cyclotides, XXC) GLP AP01141 Ref,Cryptdin-6 (Crp6, LRDLVCYCRARGCKGRERMNGTCRK 1440 animal defensin, alpha,GHLLYMLCCR mouse) AP01142 Ref, Rabbit kidney KPYCSCKWRCGIGEEEKGICHKFPIVT1441 defensin RK-2 (animal YVCCRRP defensin, alpha- defensin) AP01146Ref, Gallinacin 6 (Gal6, DTLACRQSHGSCSFVACRAPSVDIGTC 1442 Gal-6, avianbeta RGGKLKCCKWAPSS defensin, bird) AP01147 Ref, Gallinacin 8 (Gal8,DTVACRIQGNFCRAGACPPTFTISGQC 1443 Gal-8, avian beta HGGLLNCCAKIPAQdefensin, bird) AP01148 Ref, Gallinacin 3 (Gal3,IATQCRIRGGFCRVGSCRFPHIAIGKCA 1444 Gal-3, avian beta TFISCCGRAY defensin,bird) AP01152 Ref, Lactococcin Q SIWGDIGQGVGKAAYWVGKAMGNM 1445 (classIIb bacteriocin, SDVNQASRINRKKKH bacteria, chain a. For chain b, seeInfo) AP01155 Ref, Enterocin 1071 ESVFSKIGNAVGPAAYWILKGLGNMS 1446(Ent1071A, class IIb DVNQADRINRKKH bacteriocin, bacteria; chain B isEnterocin 1071B or Ent1071B, see info) AP01156 Ref, Plantaricin S (chainNKLAYNMGHYAGKATIFGLAAWALLA 1447 a, class IIb bacteriocin, bacteria)AP01159 Ref, Hinnavin II (Hin II, KWKIFKKIEHMGQNIRDGLIKAGPAV 1448 XXA,insect) QVVGQAATIYK AP01160 Ref, NK-2 (synthetic,KILRGLCKKIMRSFLRRISWDILTGKK 1449 XXA) AP01167 Ref, Plantaricin NC8LTTKLWSSWGYYLGKKARWNLKHPY 1450 (PLNC8, chain a, class VQF IIbbacteriocin, bacteria. For chain b, see Info) AP01168 Ref, Carnocyclin A(a LVAYGIAQGTAEKVVSLINAGLTVGSI 1451 circular bacteriocin,ISILGGVTVGLSGVFTAVKAAIAKQGI XXC, bacteria) KKAIQL AP01169 Ref, LactacinF (LafX, NRWGDTVLSAASGAGTGIKACKSFGP 1452 class IIb bacteriocin,WGMAICGVGGAAIGGYFGYTHN bacteria. For LafA, see Info) AP01170 Ref,Brochocin C YSSKDCLKDIGKGIGAGTVAGAAGGG 1453 (BrcC, chain BrcA,LAAGLGAIPGAFVGAHFGVIGGSAACI class IIb bacteriocin, GGLLGN bacteria. ForBrcB, see Info) AP01171 Ref, Thermophilin 13 YSGKDCLKDMGGYALAGAGSGALW1454 (chain a ThmA, 2-chain GAPAGGVGALPGAFVGAHVGAIAGG class IIbbacteriocin, FACMGGMIGNKFN bacteria. For chain B ThmB, see Info) AP01172Ref, ABP-118 (chain a: KRGPNCVGNFLGGLFAGAAAGVPLGP 1455 Abp118alpha,class IIb AGIVGGANLGMVGGALTCL bacteriocin, bacteria. For chain b:Abp118beta, see Info) AP01173 Ref, Salivaricin P (chainKRGPNCVGNFLGGLFAGAAAGVPLGP 1456 a: Sln1; class IIb AGIVGGANLGMVGGALTCLbacteriocin, bacteria. For chain b: Sln2, see Info) AP01174 Ref, MutacinIV (chain KVSGGEAVAAIGICATASAAIGGLAG 1457 a: NlmA, class IIbATLVTPYCVGTWGLIRSH bacteriocin, bacteria. For chain b: NLmB, see Info)AP01175 Ref, Lactocin 705 GMSGYIQGIPDFLKGYLHGISAANKH 1458 (chain a:Lac705alpha; KKGRLGY class IIb bacteriocin, bacteria. For chain b:Lac705beta, see Info) AP01176 Ref, Cytolysin (CylLS,TTPACFTIGLGVGALFSAKFC 1459 bacteria; Chain B: CylLL) AP01177 Ref,Plantaricin EF FNRGGYNFGKSVRHVVDAIGSVAGIL 1460 (chain a: PlnE, class IIbKSIR bacteriocin, bacteria. Chain b: PlnF) AP01178 Ref, Plantaricin JKGAWKNFWSSLRKGFYDGEAGRAIRR 1461 (chain a: PlnJ; class IIb bacteriocin,bacteria. Chain b: PlnK) AP01179 Ref, Enterocin SE-K4NGVYCNKQKCWVDWSRARSEIIDRG 1462 (class IIa bacteriocin,VKAYVNGFTKVLGGIGGR bacteria) AP01180 Ref, Acidocin J1132NPKVAHCASQIGRSTAWGAVSGA 1463 (class IIb bacteriocin, bacteria) AP01181Ref, Curvaticin L442 AYPGNGVHCGKYSCTVDKQTAIGNIG 1464 (class IIabacteriocin, NNAA bacteria) AP01182 Ref, Bacteriocin 32FTPSVSFSQNGGVVEAAAQRGYIYKK 1465 (Bac 32, class IIaYPKGAKVPNKVKMLVNIRGKQTMRT bacteriocin, bacteria) CYLMSWTASSRTAKYYYYIAP01183 Ref, Bacteriocin 43 ATYYGNGLYCNKEKCWVDWNQAKG 1466 (Bac 43,bacteriocin, EIGKIIVNGWVNHGPWAPRR bacteria) AP01184 Ref, Bacteriocin T8ATYYGNGLYCNKEKCWVDWNQAKG 1467 (Bac T8, class IIa EIGKIIVNGWVNHGPWAPRRbacteriocin, bacteria) AP01185 Ref, Enterocin B (EntB,ENDHRMPNNLNRPNNLSKGGAKCGA 1468 bacteriocin, bacteria)AIAGGLFGIPKGPLAWAAGLANVYSK CN AP01186 Ref, Acidocin AKTYYGTNGVHCTKKSLWGKVRLKNV 1469 (bacteriocin, bacteria)IPGTLCRKQSLPIKQDLKILLGWATGA FGKTFH AP01187 Ref, Enterocin Q (EntQ,MNFLKNGIAKWMTGAELQAYKKKY 1470 class IIc bacteriocin, GCLPWEKISCleaderless, i.e. no signal peptide, bacteria) AP01188 Ref, EnterocinEJ97 MLAKIKAMIKKFPNPYTLAAKLTTYEI 1471 (EntEJ97, class IIcNWYKQQYGRYPWERPVA bacteriocin, leaderless, i.e. no signal peptide,bacteria) AP01189 Ref, Enterocin RJ-11 APAGLVAKFGRPIVKKYYKQIMQFIG 1472(EntRJ-11, class IIc EGSAINKIIPWIARMWRT bacteriocin, leaderless, i.e. nosignal sequence, bacteria) AP01190 Ref, Enterocin L50 (oldMGAIAKLVAKFGWPIVKKYYKQIMQ 1473 name: pediocin L50, FIGEGWAINKIIEWIKKHIEntL50A, a two-chain class IIc bacteriocin, leaderless, i.e. no signalpeptide, bacteria. The sequence of EntL50B is provided in Info) AP01191Ref, MR10 (MR10A, MGAIAKLVAKFGWPIVKKYYKQIMQ 1474 class IIc bacteriocin,FIGEGWAINKIIDWIKKHI leaderless, i.e. no signal peptide, bacteria. Forthe sequence of chain b, see Info) AP01192 Ref, Halocin S8 (HalS8,SDCNINSNTAADVILCFNQVGSCALCS 1475 microhalocin, PTLVGGPVP archaeocins,archeae) AP01193 Ref, Halocin C8 DIDITGCSACKYAAGQVCTIGCSAAG 1476 (HalC8,microhalocins, GFICGLLGITIPVAGLSCLGFVEIVCTV archaeocins, archaea)ADEYSGCGDAVAKEACNRAGLC AP01194 Ref, Lacticin 3147CSTNTFSLSDYWGNNGAWCTLTHEC 1477 (chain A1, a two-chain MAWCK lantibiotic,bacteriocin, bacteria. The sequence of chain A2 is given in Info; XXD3)AP01195 Ref, Salivaricin A KRGSGWIATITDDCPNSVFVCC 1478 (SalA,lantibiotic, bacteriocin, bacteria) AP01196 Ref, Microcin E492ATYYGNGLYCNKEKCWVDWNQAKG 1479 (MccE492, class IIb EIGKIIVNGWVNHGPWAPRRmicrocins, bacteriocin, bacteria; BBM; u- MccE492, siderophore peptide,BBI, XXG) AP01197 Ref, Hiracin JM79 ATYYGNGLYCNKEKCWVDWNQAKG 1480(HirJM79, a Sec- EIGKIIVNGWVNHGPWAPRR dependent class II bacteriocin,bacteria) AP01198 Ref, Thermophilin 9 LSCDEGMLAVGGLGAVGGPWGAAV 1481(BlpDst, class IIb GVLVGAALYCF bacteriocin, bacteria. beta-chains:BlpUst, BlpEst, BapFst) AP01199 Ref, Penocin A (PenA,KYYGNGVHCGKKTCYVDWGQATASI 1482 class IIa bacteriocin, GKIIVNGWTQHGPWAHRbacteria) AP01200 Ref, Salivaricin B GGGVIQTISHECRMNSWQFLFTCCS 1483(SalB, lantibotic, bacteriocin, bacteria) AP01201 Ref, Lacticin 481KGGSGVIHTISHECNMNSWQFVFTCCS 1484 (lantibiotic, class I bacteriocin,bacteria) AP01202 Ref, Bacteriocin J46 KGGSGVIHTISHEVIYNSWNFVFTCCS 1485(BacJ46, bacteriocin, bacteria) AP01203 Ref, Nukacin A (NucA,KKKSGVIPTVSHDCHMNSFQFVFTCCS 1486 Nukacin ISK-1, NukISK-1, bacteriocin,bacteria) AP01204 Ref, Streptococcin A- GKNGVFKTISHECHLNTWAFLATCCS 1487FF22 (LANTIBIOTIC, class I bacteriocin, bacteria) AP01210 Ref,Jelleine-I PFKLSLHL 1488 (honeybees, insect, XXA) AP01211 Ref,Jelleine-II TPFKLSLHL 1489 (honeybees, insect, XXA) AP01212 Ref,Jelleine-III EPFKLSLHL 1490 (honeybees, insect, XXA) AP01213 Ref,Hymenoptaecin EFRGSIVIQGTKEGKSRPSLDIDYKQR 1491 (honeybees, insectVYDKNGMTGDAYGGLNIRPGQPSRQ defensin, XXcooh) HAGFEFGKEYKNGFIKGQSEVQRGPGGRLSPYFGINGGFRF AP01216 Ref, Ascaphin-1 (frog, GFRDVLKGAAKAFVKTVAGHIAN1492 XXA) AP01218 Ref, Ascaphin-3 (frog) GFRDVLKGAAKAFVKTVAGHIANI 1493AP01220 Ref, Ascaphin-5 (frog) GIKDWIKGAAKKLIKTVASNIANQ 1494 AP01222Ref, Ascaphin-7 (frog) GFKDWIKGAAKKLIKTVASSIANQ 1495 AP01223 Ref,Ascaphin-8 (frog, GFKDLLKGAAKALVKTVLF 1496 XXA) AP01226 Ref, Microcin C7MRTGNAD 1497 (MccC7, microcin C51, MccC51, class I microcins,bacteriocins, bacteria. Others: MccA; XXamp; BBPe) AP01227 Ref, MicrocinB17 VGIGGGGGGGGGGSCGGQGGGCGGC 1498 (MccB17, class I SNGCSGGNGGSGGSGSHImicrocins, bacteriocins, Gram-negative bacteria; BBPe) AP01228 Ref,Microcin V (MccV, ASGRDIAMAIGTLSGQFVAGGIGAAA 1499 (old name) Colicin V,GGVAGGAIYDYASTHKPNPAMSPSGL ColV; class II GGTIKQKPEGIPSEAWNYAAGRLCNWmicrocins, bacteriocins, SPNNLSDVCL Gram-negative bacteria) AP01229 Ref,Microcin L (MccL, GDVNWVDVGKTVATNGAGVIGGAFG 1500 class IIa microcins,AGLCGPVCAGAFAVGSSAAVAALYD bacteriocins, Gram- AAGNSNSAKQKPEGLPPEAWNYAEGnegative bacteria) RMCNWSPNNLSDVCL AP01230 Ref, Microcin MDGNDGQAELIAIGSLAGTFISPGFGSIA 1501 (MccM, class IIbGAYIGDKVHSWATTATVSPSMSPSGI microcins, bacteriocins,GLSSQFGSGRGTSSASSSAGSGS Gram-negative bacteria) AP01231 Ref, MicrocinH47 GGAPATSANAAGAAAIVGALAGIPGG 1502 (MccH47, class IIbPLGVVVGAVSAGLTTGIGSTVGSGSA microcins, bacteriocins, SSSAGGGSGram-negative bacteria) AP01232 Ref, Microcin I47MNLNGLPASTNVIDLRGKDMGTYIDA 1503 (MccI47, class IIbNGACWAPDTPSIIMYPGGSGPSYSMSS microcins, bacteriocins, STSSANSGSGram-negative bacteria) Aibellin *Ac U A U A U A Q U F U G U U P V U1504 U E E [NHC(CH2Ph)HCH2NHCH2CH2]OH Alamethicin_F-30 *Ac U P U A U A QU V U G L U P V U 1505 U E Q F OH Alamethicin_F-50 *Ac U P U A U A Q U VU G L U P V U 1506 U Q Q F OH Alamethicin_II *Ac U P U A U U Q U V U G LU P V U 1507 U E Q F OH Ampullosporin *Ac W A U U L U Q U U U Q L U Q L1508 OH Ampullosporin_B *Ac W A U U L U Q A U U Q L U Q L 1509 OHAmpullosporin_C *Ac W A U U L U Q U A U Q L U Q L 1510 OHAmpullosporin_D *Ac W A U U L U Q U U A Q L U Q L 1511 OHAmpullosporin_E1 *Ac W A U U L U Q A U U Q L A Q L 1512 OHAmpullosporin_E2 *Ac W A U U L U Q U A A Q L U Q L 1513 OHAmpullosporin_E3 *Ac W A U U L U Q U U A Q L A Q L 1514 OHAmpullosporin_E4 *Ac W A U U L U Q A A U Q L U Q L 1515 OH Antiamoebin_I*Ac F U U U J G L U U O Q J O U P F 1516 OH Antiamoebin_II *Ac F U U U JG L U U O Q J P U P F 1517 OH Antiamoebin_III *Ac F U U U U G L U U O QJ O U P F 1518 OH Antiamoebin_IV *Ac F U U U J G L U U O Q J O U P F1519 OH Antiamoebin_V *Ac F U U U J A L U U O Q J O U P F 1520 OHAntiamoebin_VI *Ac F U U U U G L U U O Q U O U P F 1521 OHAntiamoebin_VII *Ac F A U J U G L U U O Q J O U P F 1522 OHAntiamoebin_VIII *Ac F U U U J G L U U O Q U O U P F 1523 OHAntiamoebin_IX *Ac F U A U J G L U U O Q J O U P F 1524 OH Antiamoebin_X*Ac F U U U J G L J U O Q U O U P F 1525 OH Antiamoebin_XI *Ac F U U U UA L U U O Q J O U P F 1526 OH Antiamoebin_XII *Ac F U U U U G L A U O QJ O U P F 1527 OH Antiamoebin_XIII *Ac V U U U U G L U U O Q J O U P F1528 OH Antiamoebin_XIV *Ac V U U U V G L U U O Q J O U P F 1529 OHAntiamoebin_XV *Ac L U U U U G L U U O Q J O U P F 1530 OHAntiamoebin_XVI *Ac L U U U J G L U U O Q J O U P F 1531 OHAtroviridin_A *Ac U P U A U A Q U V U G L U P V U 1532 U Q Q F OHAtroviridin_B *Ac U P U A U A Q U V U G L U P V U 1533 J Q Q F OHAtroviridin_C *Ac U P U A U U Q U V U G L U P V U 1534 J Q Q F OHBergofungin_A *Ac V U U U V G L U U O Q J O U F 1535 OH Bergofungin_B*Ac V U U U V G L V U O Q U O U F 1536 OH Bergofungin_C *Ac V U U U V GL U U O Q U O U F 1537 OH Bergofungin_D *Ac V U U V G L U U O Q U O U FOH 1538 Boletusin *Ac F U A U J L Q G U U A A U P U U 1539 U Q W OHCephaibol_A *Ac F U U U U G L J U O Q J O U P F 1540 OH Cephaibol_A2 *AcF U U U U A L J U O Q J O U P F 1541 OH Cephaibol_B *Ac F U U U J G L JU O Q J O U P F 1542 OH Cephaibol_C *Ac F U U U U G L J U O Q U O U P F1543 OH Cephaibol_D *Ac F U U U U G L U U O Q U O U P F 1544 OHCephaibol_E *Ac F U U U U G L U U O Q J O U P F 1545 OH Cephaibol_P *AcF J Q U I T U L U O Q U O U P F S 1546 OH Cephaibol_Q *Ac F J Q U I T UL U P Q U O U P F S 1547 OH Cervinin_1 *Ac L U P U L U P A U P V L OH1548 Cervinin_2 *Ac L U P U L U P A U P V L OCOCH3 1549 Chrysospermin_A*Ac F U S U U L Q G U U A A U P U U 1550 U Q W OH Chrysospermin_B *Ac FU S U U L Q G U U A A U P J U U 1551 Q W OH Chrysospermin_C *Ac F U S UJ L Q G U U A A U P U U U 1552 Q W OH Chrysospermin_D *Ac F U S U J L QG U U A A U P J U U 1553 Q W OH Clonostachin *Ac U O L J O L J O U J U OJ I 1554 O[CH(CH(OH)CH2OH)CH(OH)CH(OH)CH2]OH Emerimicin_II_A *Ac W I Q UI T U L U O Q U O U P F 1555 OH Emerimicin_II_B *Ac W I Q J I T U L U OQ U O U P F 1556 OH Emerimicin_III *Ac F U U U V G L U U O Q J O U F OH1557 Emerimicin_IV *Ac F U U U V G L U U O Q J O A F OH 1558Harzianin_HB_I *Ac U N L I U P J L U P L OH 1559 Harzianin_HC_I *Ac U NL U P S V U P U L U P L OH 1560 Harzianin_HC_III *Ac U N L U P S V U P JL U P L OH 1561 Harzianin_HC_IX *Ac U N L U P A I U P J L U P L OH 1562Harzianin_HC_VI *Ac U N L U P A V U P U L U P L OH 1563Harzianin_HC_VIII *Ac U N L U P A V U P J L U P L OH 1564Harzianin_HC_VIII *Ac U N L U P A V U P J L U P L OH 1565 Harzianin_HC_X*Ac U Q L U P A V U P J L U P L OH 1566 Harzianin_HC_XI *Ac U N L U P SI U P U L U P L OH 1567 Harzianin_HC_XII *Ac U N L U P S I U P J L U P LOH 1568 Harzianin_HC_XIII *Ac U Q L U P S I U P J L U P L OH 1569Harzianin_HC_XIV *Ac U N L U P A I U P U L U P L OH 1570 Harzianin_HC_XV*Ac U Q L U P A I U P J L U P L OH 1571 Harzianin_HK_VI *Ac U N I I U PL L U P L OH 1572 Harzianin_PCU4 *Ac U N L U P S I U P U L U P V OH 1573Helioferin_A *Fa P ZZ A U I I U U AAE 1574 Helioferin_B *Fa P ZZ A U I IU U AMAE 1575 Heptaibin *Ac F U U U V G L U U O Q U O U F 1576 OHHypelcin_A *Ac U P U A U U Q L U G U U U P V U 1577 U Q Q L OHHypelcin_A_I *Ac U P U A U U Q U L U G U U P V U 1578 U Q Q L OHHypelcin_A_II *Ac U P U A U A Q U L U G U U P V U 1579 U Q Q L OHHypelcin_A_III *Ac U P U A U U Q U L U G U U P V U 1580 U Q Q [C7H16NO]Hypelcin_A_IV *Ac U P U A U U Q U I U G U U P V U 1581 U Q Q L OHHypelcin_A-III *Ac U P U A U U Q U L U G U U P V U 1582 J Q Q L OHHypelcin_A-IX *Ac U P U A U U Q U I U G U U P V U J 1583 Q Q L OHHypelcin_A-V *Ac U P U A U U Q U L U G U U P V U 1584 U Q Q I OHHypelcin_A-VI *Ac U P U A U A Q U L U G U U P V U 1585 U Q Q I OHHypelcin_A-VII *Ac U P U A U A Q U L U G U U P V U 1586 J Q Q L OHHypelcin_A-VIII *Ac U P U A U A Q U I U G U U P V U 1587 U Q Q L OHHypelcin_B_I *Ac U P U A U U Q U L U G U U P V U 1588 U E Q L OHHypelcin_B_II *Ac U P U A U A Q U L U G U U P V U 1589 U E Q L OHHypelcin_B_III *Ac U P U A U U Q U L U G U U P V U 1590 J E Q L OHHypelcin_B_IV *Ac U P U A U U Q U I U G U U P V U 1591 U E Q L OHHypelcin_B_V *Ac U P U A U U Q U L U G U U P V U 1592 U E Q I OHHypomurocin_A_I *Ac U Q V V U P L L U P L OH 1593 Hypomurocin_A_II *Ac JQ V V U P L L U P L OH 1594 Hypomurocin_A_III *Ac U Q V L U P L I U P LOH 1595 Hypomurocin_A_IV *Ac U Q I V U P L L U P L OH 1596Hypomurocin_A_V *Ac U Q I I U P L L U P L OH 1597 Hypomurocin_A_Va *Ac UQ I L U P L I U P L OH 1598 Hypomurocin_B_I *Ac U S A L U Q U V U G U UP L U U 1599 Q V OH Hypomurocin_B_II *Ac U S A L U Q U V U G U U P L U U1600 Q L OH Hypomurocin_B_IIIa *Ac U A A L U Q U V U G U U P L U U 1601Q V OH Hypomurocin_B_IIIb *Ac U S A L U Q J V U G U U P L U U 1602 Q VOH Hypomurocin_B_IV *Ac U S A L U Q U V U G J U P L U U 1603 Q V OHHypomurocin_B_V *Ac U S A L U Q U V U G J U P L U U 1604 Q L OHLeu1_Zervamicin *Ac L I Q J I T U L U O Q U O U P F OH 1605Longibrachin_A_I *Ac U A U A U A Q U V U G L U P V U 1606 U Q Q F OHLongibrachin_A_II *Ac U A U A U A Q U V U G L U P V U 1607 J Q Q F OHLongibrachin_A_III *Ac U A U A U U Q U V U G L U P V U 1608 U Q Q F OHLongibrachin_A_IV *Ac U A U A U U Q U V U G L U P V U 1609 J Q Q F OHLongibrachin_B_II *Ac U A U A U A Q U V U G L U P V U 1610 U E Q F OHLongibrachin_B_III *Ac U A U A U A Q U V U G L U P V U 1611 J E Q F OHLP237_F5 *Oc U P Y U Q Q U Zor Q A L OH 1612 LP237_F7 *Ac U P F U Q Q UU Q A L OH 1613 LP237_F8 *Oc U P F U Q Q U Zor Q A L OH 1614 NA_VII *AcU A A U J Q U U U S L U OCH3 1615 Paracelsin_A *Ac U A U A U A Q U V U GU U P V U 1616 U Q Q F OH Paracelsin_B *Ac U A U A U A Q U L U G U U P VU 1617 U Q Q F OH Paracelsin_C *Ac U A U A U U Q U V U G U U P V U 1618U Q Q F OH Paracelsin_D *Ac U A U A U U Q U L U G U U P V U 1619 U Q Q FOH Paracelsin_E *Ac U A U A U A Q U L U G U A P V U 1620 U Q Q F OHPeptaibolin *Ac L U L U F OH 1621 Peptaivirin_A *Ac F U A U J L Q G U UA A U P J U U 1622 Q W OH Peptaivirin_B *Ac F U S U J L Q G U U A A U PJ U U 1623 Q F OH Polysporin_A *Ac U P U A U U Q U V U G V U P V U 1624U Q Q F OH Polysporin_B *Ac U P U A U U Q U V U G L U P V U 1625 U Q Q FOH Polysporin_C *Ac U P U A U U Q U I U G L U P V U 1626 U Q Q F OHPolysporin_D *Ac U P U A U U Q U I U G L U P V U 1627 V Q Q F OHPseudokinin_KLIII *Ac U N I I U P L L U P NH2 1628 Pseudokinin_KLVI *AcU N I I U P L V 1629 hydroxyketopiperazine Samarosporin_I *Ac F U U U VG L U U O Q J O A F OH 1630 Samarosporin_II *Ac F U U U V G L U U O Q JO U F OH 1631 Saturnisporin_SA_I *Ac U A U A U A Q U L U G U U P V U1632 U Q Q F OH Saturnisporin_SA_II *Ac U A U A U A Q U L U G U U P V U1633 J Q Q F OH Saturnisporin_SA_III *Ac U A U A U U Q U L U G U U P V U1634 U Q Q F OH Saturnisporin_SA_IV *Ac U A U A U U Q U L U G U U P V U1635 J Q Q F OH Stilbellin_I *Ac F U U U V G L U U O Q J O A F OH 1636Stilbellin_II *Ac F U U U V G L U U O Q J O U F OH 1637 Stilboflavin_A_1*Ac U P U A U A Q U V U G U U P V U 1638 U E Q V OH Stilboflavin_A_2 *AcU P U A U A Q U L U G U U P V U 1639 U E Q V OH Stilboflavin_A_3 *Ac U PU A U U Q U V U G U A P V U 1640 U E Q L OH Stilboflavin_A_4 *Ac U P U AU A Q U L U G U U P V U 1641 U E Q L OH Stilboflavin_A_5 *Ac U P U A U UQ U L U G U U P V U 1642 U E Q V OH Stilboflavin_A_6 *Ac U P U A U A Q UL U G U U P V U 1643 U E Q J OH Stilboflavin_A_7 *Ac U P U A U U Q U L UG U U P V U 1644 U E Q I OH Stilboflavin_B_1 *Ac U P U A U A Q U V U G UU P V U 1645 U Q Q V OH Stilboflavin_B_2 *Ac U P U A U A Q U L U G U U PV U 1646 U Q Q V OH Stilboflavin_B_3 *Ac U P U A U A Q U V U G U U P V U1647 U Q Q L OH Stilboflavin_B_4 *Ac U P U A U A Q U L U G U U P V U1648 U Q Q L OH Stilboflavin_B_5 *Ac U P U A U U Q U L U G U U P V U1649 U Q Q V OH Stilboflavin_B_6 *Ac U P U A U U Q U V U G U U P V U1650 U Q Q V OH Stilboflavin_B_7 *Ac U P U A U U Q U L U G U U P V U1651 U Q Q L OH Stilboflavin_B_8 *Ac U P U A U U Q U V U G U U P V U1652 U Q Q L OH Stilboflavin_B_9 *Ac U P U A U U Q U L U G U U P V U1653 U Q Q I OH Stilboflavin_B_10 *Ac U P U A U U Q U V U G U U P V U1654 U Q Q I OH Suzukacillin *Ac U A U A U A Q U U U G L U P V U 1655 UQ Q F OH Trichobrachin_A-I *Ac U N L L U P L U U P L OH 1656Trichobrachin_A-II *Ac U N L L U P V L U P V OH 1657 Trichobrachin_A-III*Ac U N V L U P L L U P V OH 1658 Trichobrachin_A-IV *Ac U N L V U P L LU P V OH 1659 Trichobrachin_B-I *Ac U N L L U P V U V P L OH 1660Trichobrachin_B-II *Ac U N V L U P L U V P L OH 1661 Trichobrachin_B-III*Ac U N L V U P L U V P L OH 1662 Trichobrachin_B-IV *Ac U N L L U P L UV P V OH 1663 Trichocellin_TC-A-I *Ac U A U A U A Q U L U G U U P V U1664 U Q Q F OH Trichocellin_TC-A-II *Ac U A U A U A Q U L U G U U P V U1665 J Q Q F OH Trichocellin_TC-A-III *Ac U A U A U A Q U I U G U U P VU 1666 U Q Q F OH Trichocellin_TC-A-IV *Ac U A U A U A Q U I U G U U P VU 1667 J Q Q F OH Trichocellin_TC-A-V *Ac U A U A U A Q U L U G L U P VU 1668 U Q Q F OH Trichocellin_TC-A-VI *Ac U A U A U A Q U L U G L U P VU 1669 J Q Q F OH Trichocellin_TC-A-VII *Ac U A U A U A Q U I U G L U PV U 1670 U Q Q F OH Trichocellin_TC-A-VIII *Ac U A U A U A Q U I U G L UP V U J 1671 Q Q F OH Trichocellin_TC-B-I *Ac U A U A U A Q U L U G U UP V U 1672 U E Q F OH Trichocellin_TC-B-II *Ac U A U A U A Q U L U G U UP V U 1673 J E Q F OH Trichodecenin_TD_I *(Z)-4-decenoyl G G L U G I LOH 1674 Trichodecenin_TD_II *(Z)-4-decenoyl G G L U G L L OH 1675Trichogin_A_IV *Oc U G L U G G L U G I L OH 1676 Trichokindin_Ia *Ac U SA U U Q J L U A U U P L U U 1677 Q I OH Trichokindin_Ib *Ac U S A U J QU L U A U U P L U U 1678 Q I OH Trichokindin_IIa *Ac U S A U U Q U L U AJ U P L U U 1679 Q I OH Trichokindin_IIb *Ac U S A U J Q J L U A U U P LU U Q 1680 L OH Trichokindin_IIIa *Ac U S A U U Q J L U A J U P L U U Q1681 L OH Trichokindin_IIIb *Ac U S A U J Q U L U A J U P L U U Q 1682 LOH Trichokindin_IV *Ac U S A U J Q J L U A U U P L U U Q 1683 I OHTrichokindin_Va *Ac U S A U U Q J L U A J U P L U U Q 1684 I OHTrichokindin_Vb *Ac U S A U J Q U L U A J U P L U U Q 1685 I OHTrichokindin_VI *Ac U S A U J Q J L U A J U P L U U Q 1686 L OHTrichokindin_VII *Ac U S A U J Q J L U A J U P L U U Q I 1687 OHTrichokonin_Ia *Ac U A U A U A Q U V U G L A P V U 1688 U Q Q F OHTrichokonin_Ib *Ac U G U A U A Q U V U G L U P V U 1689 U Q Q F OHTrichokonin_IIa *Ac U A U A U A Q U V U G L U P A U 1690 U Q Q F OHTrichokonin_IIb *Ac A A U A U A Q U V U G L U P V U 1691 U Q Q F OHTrichokonin_IIc *Ac U A A A U A Q U V U G L U P V U 1692 U Q Q F OHTrichokonin_V *Ac U A U A U Q U V U G L U P V U U 1693 Q Q F OHTrichokonin_VII *Ac U A U A U A Q U V U G L U P V U 1694 J Q Q F OHTrichokonin_VIII *Ac U A U A U U Q U V U G L U P V U 1695 U Q Q F OHTrichokonin_IX *Ac U A U A U A Q U V U G L U P V U 1696 J Q Q F OHTricholongin_BI *Ac U G F U U Q U U U S L U P V U U 1697 Q Q L OHTricholongin_BII *Ac U G F U U Q U U U S L U P V U J Q 1698 Q L OHTrichopolyn_I *Fa P ZZ A U U I A U U AMAE 1699 Trichopolyn_II *Fa P ZZ AU U V A U U AMAE 1700 Trichopolyn_III *Fa P ZZ A U U I A U A AMAE 1701Trichopolyn_IV *Fa P ZZ A U U V A U A AMAE 1702 Trichopolyn_V *Fa′ P ZZA U U I A U U AMAE 1703 Trichorovin_TV_Ia *Ac U N V Lx U P Lx Lx U P VOH 1704 Trichorovin_TV_Ib *Ac U N V V U P Lx Lx U P Lx OH 1705Trichorovin_TV_IIa *Ac U N V V U P Lx Lx U P Lx OH 1706Trichorovin_TV_IIb *Ac U N Lx V U P Lx Lx U P V OH 1707Trichorovin_TV_IIIa *Ac U Q V V U P Lx Lx U P Lx OH 1708Trichorovin_TV_IIIb *Ac U Q V Lx U P Lx Lx U P V OH 1709Trichorovin_TV_IVa *Ac U Q V V U P Lx Lx U P Lx OH 1710Trichorovin_TV_IVb *Ac U Q Lx V U P Lx Lx U P V OH 1711Trichorovin_TV_IVc *Ac U N V Lx U P Lx Lx U P Lx OH 1712Trichorovin_TV_IXa *Ac U Q V Lx U P Lx Lx U P Lx OH 1713Trichorovin_TV_IXb *Ac U Q Lx Lx U P Lx Lx U P V OH 1714Trichorovin_TV_Va *Ac U N V Lx U P Lx Lx U P Lx OH 1715Trichorovin_TV_Vb *Ac U N Lx Lx U P Lx Lx U P V OH 1716Trichorovin_TV_VIa *Ac U N V Lx U P Lx Lx U P Lx OH 1717Trichorovin_TV_VIb *Ac U N Lx Lx U P Lx Lx U P V OH 1718Trichorovin_TV_VIIa *Ac U N Lx V U P Lx Lx U P Lx OH 1719Trichorovin_TV_VIIb *Ac U Q V Lx U P Lx Lx U P V OH 1720Trichorovin_TV_VIII *Ac U Q V Lx U P Lx Lx U P Lx OH 1721Trichorovin_TV_Xa *Ac U Q Lx V U P Lx Lx U P Lx OH 1722Trichorovin_TV_Xb *Ac U N Lx Lx U P Lx Lx U P Lx OH 1723Trichorovin_TV_XIIa *Ac U N I I U P L L U P I OH 1724Trichorovin_TV_XIIb *Ac U N Lx Lx U P Lx Lx U P L OH 1725Trichorovin_TV_XIII *Ac U Q Lx Lx U P Lx Lx U P Lx OH 1726Trichorovin_TV_XIV *Ac U Q Lx Lx U P Lx Lx U P Lx OH 1727 Trichorozin_I*Ac U N I L U P I L U P V OH 1728 Trichorozin_II *Ac U Q I L U P I L U PV OH 1729 Trichorozin_III *Ac U N I L U P I L U P L OH 1730Trichorozin_IV *Ac U Q I L U P I L U P L OH 1731 Trichorzianine_TA_IIIc*Ac U A A U U Q U U U S L U P V U I 1732 Q Q W OH Trichorzianine_TB_IIa*Ac U A A U U Q U U U S L U P L U I Q 1733 E W OH Trichorzianine_TB_IIIc*Ac U A A U U Q U U U S L U P V U I 1734 Q E W OH Trichorzianine_TB_IVb*Ac U A A U J Q U U U S L U P V U I Q 1735 E W OH Trichorzianine_TB_Vb*Ac U A A U U Q U U U S L U P L U I Q 1736 E F OH Trichorzianine_TB_VIa*Ac U A A U J Q U U U S L U P L U I Q 1737 E F OH Trichorzianine_TB_VIb*Ac U A A U U Q U U U S L U P V U I 1738 Q E F OH Trichorzianine_TB_VII*Ac U A A U J Q U U U S L U P V U I Q 1739 E F OH Trichorzin_HA_I *Ac UG A U U Q U V U G L U P L U U 1740 Q L OH Trichorzin_HA_II *Ac U G A U UQ U V U G L U P L U J 1741 Q L OH Trichorzin_HA_III *Ac U G A U J Q U VU G L U P L U U 1742 Q L OH Trichorzin_HA_V *Ac U G A U J Q U V U G L UP L U J Q 1743 L OH Trichorzin_HA_VI *Ac U G A U J Q J V U G L U P L U JQ 1744 L OH Trichorzin_HA_VII *Ac U G A U J Q V V U G L U P L U J Q 1745L OH Trichorzin_MA_I *Ac U S A U U Q U L U G L U P L U U 1746 Q V OHTrichorzin_MA_II *Ac U S A U J Q U L U G L U P L U U Q 1747 V OHTrichorzin_MA_III *Ac U S A U J Q J L U G L U P L U U Q 1748 V OHTrichorzin_PA_II *Ac U S A U J Q U V U G L U P L U U 1749 Q W OHTrichorzin_PA_IV *Ac U S A U J Q J V U G L U P L U U Q 1750 W OHTrichorzin_PA_V *Ac U S A J J Q U V U G L U P L U U Q 1751 W OHTrichorzin_PA_VI *Ac U S A U J Q U V U G L U P L U U 1752 Q F OHTrichorzin_PA_VII *Ac U S A J J Q U V U G L U P L U U Q 1753 W OHTrichorzin_PA_VIII *Ac U S A U J Q J V U G L U P L U U Q 1754 F OHTrichorzin_PA_IX *Ac U S A J J Q U V U G L U P L U U Q 1755 F OHTrichorzin_PAU4 *Ac U S A U U Q U V U G L U P L U U 1756 Q W OHTrichosporin_TS-B-1a-1 *Ac U A G U A U Q U Lx A A Vx A P V 1757 U Vx Q QF OH Trichosporin_TS-B-1a-2 *Ac U A G A U U Q U Lx A A Vx A P V 1758 UVx Q Q F OH Trichosporin_TS-B-1b *Ac U A G A U U Q U Lx U G Lx A P V1759 U A Q Q F OH Trichosporin_TS-B-1d *Ac U A S A U U Q U Lx U G Lx A PV 1760 U U Q Q F OH Trichosporin_TS-B-1e *Ac U A G A U U Q U Lx U G Lx UP V 1761 U U Q Q F OH Trichosporin_TS-B-1f *Ac U A S A U U Q U Lx U G LxU P V 1762 U U Q Q F OH Trichosporin_TS-B-1g *Ac U A G A U U Q U Lx U GLx A P V 1763 U U Q Q F OH Trichosporin_TS-B-1h *Ac U A G A U U Q U Lx UG Lx U P V 1764 U Vx Q Q F OH Trichosporin_TS-B-Ia *Ac U A S A U U Q U LU G L U P V U 1765 U Q Q F OH Trichosporin_TS-B-IIIa *Ac U A A A U U Q UL U G L U P V U 1766 U Q Q F OH Trichosporin_TS-B-IIIb *Ac U A A A U U QU I U G L U P V U 1767 A Q Q F OH Trichosporin_TS-B-IIIc *Ac U A A A A UQ U I U G L U P V U 1768 U Q Q F OH Trichosporin_TS-B-IIId *Ac U A A A UU Q U V U G L U P V U 1769 U Q Q F OH Trichosporin_TS-B-IVb *Ac U A A AU U Q U L U G L U P V U 1770 J Q Q F OH Trichosporin_TS-B-IVc *Ac U A UA U U Q U V U G L U P V U 1771 U Q Q F OH Trichosporin_TS-B-IVd *Ac U AA A U U Q U V U G L U P V U 1772 J Q Q F OH Trichosporin_TS-B-V *Ac U AA A U U Q U I U G L U P V U 1773 U Q Q F OH Trichosporin_TS-B-VIa *Ac UA U A U U Q U I U G L U P V U 1774 U Q Q F OH Trichosporin_TS-B-VIb *AcU A A A U U Q U I U G L U P V U J 1775 Q Q F OH Trichotoxin_A-40 *Ac U GU L U E U U U A U U P L U J 1776 Q V OH Trichotoxin_A-40_I *Ac U G U L UQ U U A A U U P L U U 1777 E V OH Trichotoxin_A-40_II *Ac U G U L U Q UU U A A U P L U U 1778 E V OH Trichotoxin_A-40_III *Ac U G U L U Q U U AA U U P L U J 1779 E V OH Trichotoxin_A-40_IV *Ac U G U L U Q U U U A UU P L U U 1780 E V OH Trichotoxin_A-40_V *Ac U G U L U Q U U U A U U P LU J 1781 E V OH Trichotoxin_A-40_Va *Ac U A U L U Q U U U A U U P L U U1782 E V OH Trichotoxin_A-50_E *Ac U G U L U Q U U U A A U P L U U 1783Q V OH Trichotoxin_A-50_F *Ac U G U L U Q U U A A A U P L U J 1784 Q VOH Trichotoxin_A-50_G *Ac U G U L U Q U U U A A U P L U J 1785 Q V OHTrichotoxin_A-50_H *Ac U A U L U Q U U U A A U P L U J 1786 Q V OHTrichotoxin_A-50_I *Ac U G U L U Q U U U A U U P L U J 1787 Q V OHTrichotoxin_A-50_J *Ac U A U L U Q U U U A U U P L U J 1788 Q V OHTrichovirin-Ia *Ac U G A L A Q Vx V U G U U P L U 1789 U Q L OHTrichovirin-Ib *Ac U G A L U Q A V U G J U P L U U 1790 Q L OHTrichovirin-IIa *Ac U G A L A Q U V U G J U P L U U 1791 Q L OHTrichovirin-IIb *Ac U G A L U Q U V U G U U P L U U 1792 Q L OHTrichovirin-IIc *Ac U G A L U Q Vx V U G U U P L U 1793 U Q L OHTrichovirin-IIIa *Ac U G A L U Q J V U G U U P L U U 1794 Q L OHTrichovirin-IIIb *Ac U G A L J Q J U U G U U P L U U Q 1795 L OHTrichovirin-IVa *Ac U G A L J Q J V U G U U P L U U Q 1796 L OHTrichovirin-IVb *Ac U G A L U Q U V U G J U P L U U 1797 Q L OHTrichovirin-V *Ac U G A L U Q J V U G J U P L U U Q 1798 L OHTrichovirin-VIa *Ac U G A L U Q J L U G J U P L U U Q 1799 L OHTrichovirin-VIb *Ac U G A L J Q J V U G J U P L U U Q 1800 L OHTrikoningin_KA_V *Ac U G A U I Q U U U S L U P V U I Q 1801 Q L OHTrikoningin_KB_I *Oc U G V U G G V U G I L OH 1802 Trikoningin_KB_II *OcJ G V U G G V U G I L OH 1803 Tylopeptin_A *Ac W V U J A Q A U S U A L UQ L 1804 OH Tylopeptin_B *Ac W V U U A Q A U S U A L U Q L 1805 OH XR586*Ac W J Q U I T U L U P Q U O J P F G 1806 OH Zervamicin_A-1-16 *Boc W IA U I V U L U P A U P U P F 1807 OCH3 Zervamicin_ZIA *Ac W I E J V T U LU O Q U O U P F 1808 OH Zervamicin_ZIB *Ac W V E J I T U L U O Q U O U PF 1809 OH Zervamicin_ZIB′ *Ac W I E U I T U L U O Q U O U P F 1810 OHZervamicin_ZIC *Ac W I E J I T U L U O Q U O U P F 1811 OHZervamicin_ZII-1 *Ac W I Q U V T U L U O Q U O U P F 1812 OHZervamicin_ZII-2 *Ac W I Q U I T U V U O Q U O U P F 1813 OHZervamicin_ZII-3 *Ac W V Q U I T U L U O Q U O U P F 1814 OHZervamicin_ZII-4 *Ac W I Q J V T U L U O Q U O U P F 1815 OHZervamicin_ZII-5 *Ac W I Q J I T U V U O Q U O U P F 1816 OHZervamicin_ZIIA *Ac W I Q U I T U L U O Q U O U P F 1817 OHZervamicin_ZIIB *Ac W I Q J I T U L U O Q U O U P F 1818 OH CAMEL135(CAM135) GWRLIKKILRVFKGL 1819 Novispirin G2 KNLRIIRKGIHIIKKY* 1820 B-33FKKFWKWFRRF 1821 B-34 LKRFLKWFKRF 1822 B-35 KLFKRWKHLFR 1823 B-36RLLKRFKHLFK 1824 B-37 FKTFLKWLHRF 1825 B-38 IKQLLHFFQRF 1826 B-39KLLQTFKQIFR 1827 B-40 RILKELKNLFK 1828 B-41 LKQFVHFIHRF 1829 B-42VKTLLHIFQRF 1830 B-43 KLVEQLKEIFR 1831 B-44 RVLQEIKQILK 1832 B-45VKNLAELVHRF 1833 B-46 ATHLLHALQRF 1834 B-47 KLAENVKEILR 1835 B-48RALHEAKEALK 1836 B-49 FHYFWHWFHRF 1837 B-50 LYHFLHWFQRF 1838 B-51YLFQTWQHLFR 1839 B-52 YLLTEFQHLFK 1840 B-53 FKTFLQWLHRF 1841 B-54IKTLLHFFQRF 1842 B-55 KLLQTFNQIFR 1843 B-56 TILQSLKNIFK 1844 B-57LKQFVKFIHRF 1845 B-58 VKQLLKIFNRF 1846 B-59 KLVQQLKNIFR 1847 B-60RVLNQVKQILK 1848 B-61 VKKLAKLVRRF 1849 B-62 AKRLLKVLKRF 1850 B-63KLAQKVKRVLR 1851 B-64 RALKRIKHVLK 1852 1C-1 RRRRWWW 1853 1C-2 RRWWRRW1854 1C-3 RRRWWWR 1855 1C-4 RWRWRWR 1856 2C-1 RRRFWWR 1857 2C-2 RRWWRRF*1858 2C-3 RRRWWWF* 1859 2C-4 RWRWRWF* 1860 3C-1 RRRRWWK 1861 3C-2RRWWRRK 1862 3C-3 RRRWWWK 1863 3C-4 RWRWRWK 1864 4C-1 RRRKWWK 1865 4C-2RRWKRRK 1866 4C-3 RRRKWWK 1867 4C-4 RWRKRWK 1868 a-3 LHLLHQLLHLLHQF*1869 a-4 AQAAHQAAHAAHQF* 1870 a-5 KLKKLLKKLKKLLK 1871 a-6LKLLKKLLKLLKKF* 1872 a-7 LQLLKQLLKLLKQF* 1873 a-8 AQAAKQAAKAAKQF* 1874a-9 RWRRWWRHFHHFFH* 1875 a-10 KLKKLLKRWRRWWR 1876 a-11 RWRRLLKKLHHLLH*1877 a-12 KLKKLLKHLHHLLH* 1878 BD-1 FVF RHK WVW KHR FLF 1879 BD-2 VFIHRH VWV HKH VLF 1880 BD-3 WR WR AR WR WR LR WR F 1881 BD-4 WR IH LR ARLH VK FR F 1882 BD-5 LR IH AR FK VH IR LK F 1883 BD-6 FH IK FR VH LK VRFH F 1884 BD-7 FH VK IH FR LH VK FH F 1885 BD-8 LH IH AH FH VH IH LH F1886 BD-9 FK IH FR LK VH IR FK F 1887 BD-10 FK AH IR FK LR VK FH F 1888BD-11 LK AK IK FK VK LK IK F 1889 BD-12 WIW KHK FL HRH FLF 1890 BD-13VFL HRH VI KHK LVF 1891 BD-14 FL HKH VL RHR IVF 1892 BD-15 VF KHK IV HRHILF 1893 BD-16 FLF KH LFL HR IFF 1894 BD-17 LF KH ILI HR VIF 1895 BD-18FL HKH LF KHK LF 1896 BD-19 VF RHR FI HRH VF 1897 BD-20 FI HK LV HKH VLF1898 BD-21 VL RH LF RHR IVF 1899 BD-22 LV HK LIL RH LLF 1900 BD-23 VF KRVLI HK LIF 1901 BD-24 IV RK FLF RHK VF 1902 BD-25 VL KH VIA HKR LF 1903BD-26 FI RK FLF KH LF 1904 BD-27 VI RH VWV RK LF 1905 BD-28 FLF RHR FRHR LVF 1906 BD-29 LFL HKH A KHK FLF 1907 BD-30 F KHK F KHK FIF 1908BD-31 L RHR L RHR LIF 1909 BD-32 LIL K FLF K FVF 1910 BD-33 VLI R ILV RVIF 1911 BD-34 F RHR F RHR F 1912 BD-35 L KHK L KHK F 1913 BD-36 F K FKHK LIF 1914 BD-37 L R L RHR VLF 1915 BD-38 F K FLF K FLF 1916 BD-39 L RLFL R WLF 1917 BD-40 F K FLF KHK F 1918 BD-41 L R LFL RHR F 1919 BD-42 FK FLF K F 1920 BD-43 L R LFL R F 1921 AA-1 HHFFHHFHHFFHHF* 1922 AA-2FHFFHHFFHFFHHF* 1923 AA-3 KLLK-GAT-FHFFHHFFHFFHHF 1924 AA-4KLLK-FHFFHHFFHFFHHF 1925 AA-5 FHFFHHFFHFFHHFKLLK 1926 RIP YSPWTNF* 1927Abreviations: U - Aminoisobutyric Acid (Aib); J - Isovaline (Iva); O -Hydroxyproline (Hyp); Z - Ethylnorvaline (EtNor); x or xx means L or Iat that position; Ac - optionally acetylated N-term; OH, OCH3 - optionalC-term; Alkane long chains are noted in brackets; *optionally amidatedC-terminus.

A number of antimicrobial peptides are also disclosed in U.S. Pat. Nos.7,271,239, 7,223,840, 7,176,276, 6,809,181, 6,699,689, 6,420,116,6,358,921, 6,316,594, 6,235,973, 6,183,992, 6,143,498, 6,042,848,6,040,291, 5,936,063, 5,830,993, 5,428,016, 5,424,396, 5,032,574,4,623,733, which are incorporated herein by reference for the disclosureof particular antimicrobial peptides.

Ligands.

In certain embodiments the effector can comprise one or more ligands,epitope tags, and/or antibodies. In certain embodiments preferredligands and antibodies include those that bind to surface markers onimmune cells. Chimeric moieties utilizing such antibodies as effectormolecules act as bifunctional linkers establishing an associationbetween the immune cells bearing binding partner for the ligand orantibody and the target microorganism(s).

The term “epitope tag” or “affinity tag” are used interchangeablyherein, and as used refers to a molecule or domain of a molecule that isspecifically recognized by an antibody or other binding partner. Theterm also refers to the binding partner complex as well. Thus, forexample, biotin or a biotin/avidin complex are both regarded as anaffinity tag. In addition to epitopes recognized in epitope/antibodyinteractions, affinity tags also comprise “epitopes” recognized by otherbinding molecules (e.g. ligands bound by receptors), ligands bound byother ligands to form heterodimers or homodimers, His₆ bound by Ni-NTA,biotin bound by avidin, streptavidin, or anti-biotin antibodies, and thelike.

Epitope tags are well known to those of skill in the art. Moreover,antibodies specific to a wide variety of epitope tags are commerciallyavailable. These include but are not limited to antibodies against theDYKDDDDK (SEQ ID NO:1928) epitope, c-myc antibodies (available fromSigma, St. Louis), the HNK-1 carbohydrate epitope, the HA epitope, theHSV epitope, the His₄ (SEQ ID NO:1929), His₅ (SEQ ID NO:1930), and His₆(SEQ ID NO:1931) epitopes that are recognized by the His epitopespecific antibodies (see, e.g., Qiagen), and the like. In addition,vectors for epitope tagging proteins are commercially available. Thus,for example, the pCMV-Tag1 vector is an epitope tagging vector designedfor gene expression in mammalian cells. A target gene inserted into thepCMV-Tag1 vector can be tagged with the FLAG® epitope (N-terminal,C-terminal or internal tagging), the c-myc epitope (C-terminal) or boththe FLAG (N-terminal) and c-myc (C-terminal) epitopes.

Lipids and Liposomes.

In certain embodiments the effectors comprise one or more microparticlesor nanoparticles that can be loaded with an effector agent (e.g., apharmaceutical, a label, etc.). In certain embodiments themicroparticles or nanoparticles are lipidic particles. Lipidic particlesare microparticles or nanoparticles that include at least one lipidcomponent forming a condensed lipid phase. Typically, a lipidicnanoparticle has preponderance of lipids in its composition. Variouscondensed lipid phases include solid amorphous or true crystallinephases; isomorphic liquid phases (droplets); and various hydratedmesomorphic oriented lipid phases such as liquid crystalline andpseudocrystalline bilayer phases (L-alpha, L-beta, P-beta, Lc),interdigitated bilayer phases, and nonlamellar phases (see, e.g., TheStructure of Biological Membranes, ed. by P. Yeagle, CRC Press, BocaRaton, Fla., 1991). Lipidic microparticles include, but are not limitedto a liposome, a lipid-nucleic acid complex, a lipid-drug complex, alipid-label complex, a solid lipid particle, a microemulsion droplet,and the like. Methods of making and using these types of lipidicmicroparticles and nanoparticles, as well as attachment of affinitymoieties, e.g., antibodies, to them are known in the art (see, e.g.,U.S. Pat. Nos. 5,077,057; 5,100,591; 5,616,334; 6,406,713; 5,576,016;6,248,363; Bondi et al. (2003) Drug Delivery 10: 245-250; Pedersen etal., (2006) Eur. J. Pharm. Biopharm. 62: 155-162, 2006 (solid lipidparticles); U.S. Pat. Nos. 5,534,502; 6,720,001; Shiokawa et al. (2005)Clin. Cancer Res. 11: 2018-2025 (microemulsions); U.S. Pat. No.6,071,533 (lipid-nucleic acid complexes), and the like).

A liposome is generally defined as a particle comprising one or morelipid bilayers enclosing an interior, typically an aqueous interior.Thus, a liposome is often a vesicle formed by a bilayer lipid membrane.There are many methods for the preparation of liposomes. Some of themare used to prepare small vesicles (d<0.05 micrometer), some for largervesicles (d>0.05 micrometer). Some are used to prepare multilamellarvesicles, some for unilamellar ones. Methods for liposome preparationare exhaustively described in several review articles such as Szoka andPapahadjopoulos (1980) Ann. Rev. Biophys. Bioeng., 9: 467, Deamer andUster (1983) Pp. 27-51 In: Liposomes, ed. M. J. Ostro, Marcel Dekker,New York, and the like.

In various embodiments the liposomes include a surface coating of ahydrophilic polymer chain. “Surface-coating” refers to the coating ofany hydrophilic polymer on the surface of liposomes. The hydrophilicpolymer is included in the liposome by including in the liposomecomposition one or more vesicle-forming lipids derivatized with ahydrophilic polymer chain. In certain embodiments, vesicle-forminglipids with diacyl chains, such as phospholipids, are preferred. Oneillustrative phospholipid is phosphatidylethanolamine (PE), whichcontains a reactive amino group convenient for coupling to the activatedpolymers. One illustrative PE is distearoyl PE (DSPE). Another exampleis non-phospholipid double chain amphiphilic lipids, such as diacyl- ordialkylglycerols, derivatized with a hydrophilic polymer chain.

In certain embodiments a hydrophilic polymer for use in coupling to avesicle forming lipid is polyethyleneglycol (PEG), preferably as a PEGchain having a molecular weight between 1,000-10,000 Daltons, morepreferably between 1,000-5,000 Daltons, most preferably between2,000-5,000 Daltons. Methoxy or ethoxy-capped analogues of PEG are alsouseful hydrophilic polymers, commercially available in a variety ofpolymer sizes, e.g., 120-20,000 Daltons.

Other hydrophilic polymers that can be suitable include, but are notlimited to polylactic acid, polyglycolic acid, polyvinylpyrrolidone,polymethyloxazoline, polyethyloxazoline, polyhydroxypropylmethacrylamide, polymethacrylamide, polydimethylacrylamide, andderivatized celluloses, such as hydroxymethylcellulose orhydroxyethylcellulose.

Preparation of lipid-polymer conjugates containing these polymersattached to a suitable lipid, such as PE, have been described, forexample in U.S. Pat. No. 5,395,

The liposomes can, optionally be prepared for attachment to one or moretargeting moieties described herein. Here the lipid component includedin the liposomes would include either a lipid derivatized with thetargeting moiety, or a lipid having a polar-head chemical group, e.g.,on a linker, that can be derivatized with the targeting moiety inpreformed liposomes, according to known methods.

Methods of functionalizing lipids and liposomes with affinity moietiessuch as antibodies are well known to those of skill in the art (see,e.g., DE 3,218,121; Epstein et al. (1985) Proc. Natl. Acad. Sci., USA,82:3688 (1985); Hwang et al. (1980) Proc. Natl. Acad. Sci., USA, 77:4030; EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP 142,641; Japanesepatent application 83-118008; U.S. Pat. Nos. 4,485,045 and 4,544,545;and EP 102,324, all of which are incorporated herein by reference).

Polymeric Microparticles and/or Nanoparticles.

In certain embodiments the effector(s) comprise polymeric microparticlesand/or nanoparticles and/or micelles.

Microparticle and nanoparticle-based drug delivery systems haveconsiderable potential for treatment of various microorganisms.Technological advantages of polymeric microparticles or nanoparticlesused as drug carriers are high stability, high carrier capacity,feasibility of incorporation of both hydrophilic and hydrophobicsubstances, and feasibility of variable routes of administration,including oral application and inhalation. Polymeric nanoparticles canalso be designed to allow controlled (sustained) drug release from thematrix. These properties of nanoparticles enable improvement of drugbioavailability and reduction of the dosing frequency.

Polymeric nanoparticles are typically micron or submicron (<1 μm)colloidal particles. This definition includes monolithic nanoparticles(nanospheres) in which the drug is adsorbed, dissolved, or dispersedthroughout the matrix and nanocapsules in which the drug is confined toan aqueous or oily core surrounded by a shell-like wall. Alternatively,in certain embodiments, the drug can be covalently attached to thesurface or into the matrix.

Polymeric microparticles and nanoparticles are typically made frombiocompatible and biodegradable materials such as polymers, eithernatural (e.g., gelatin, albumin) or synthetic (e.g., polylactides,polyalkylcyanoacrylates), or solid lipids. In the body, the drug loadedin nanoparticles is usually released from the matrix by diffusion,swelling, erosion, or degradation. One commonly used material ispoly(lactide-co-glycolide) (PLG).

Methods of fabricating and loading polymeric nanoparticles ormicroparticles are well known to those of skill in the art. Thus, forexample, Matsumoto et al. (1999) Intl. J. Pharmaceutics, 185: 93-101teaches the fabrication of poly(L-lactide)-poly(ethyleneglycol)-poly(L-lactide) nanoparticles, Chawla et al. (2002) Intl. J.Pharmaceutics 249: 127-138, teaches the fabrication and use ofpoly(ε-caprolactone) nanoparticles delivery of tamifoxen, and Bodmeieret al. (1988) Intl. J. Pharmaceutics, 43: 179-186, teaches thepreparation of poly(D,L-lactide) microspheres using a solventevaporation method.” Intl. J. Pharmaceutics, 1988, 43, 179-186. Othernanoparticle formulations are described, for example, by Williams et al.(2003) J. Controlled Release, 91: 167-172; Leroux et al. (1996) J.Controlled Release, 39: 339-350; Soppimath et al. (2001) J. ControlledRelease, 70:1-20; Brannon-Peppas (1995) Intl. J. Pharmaceutics, 116:1-9; and the like.

Peptide Preparation.

The peptides described herein can be chemically synthesized usingstandard chemical peptide synthesis techniques or, particularly wherethe peptide does not comprise “D” amino acid residues, the peptide canbe recombinantly expressed. Where the “D” polypeptides are recombinantlyexpressed, a host organism (e.g. bacteria, plant, fungal cells, etc.)can be cultured in an environment where one or more of the amino acidsis provided to the organism exclusively in a D form. Recombinantlyexpressed peptides in such a system then incorporate those D aminoacids.

In certain embodiments, D amino acids can be incorporated inrecombinantly expressed peptides using modified amino acyl-tRNAsynthetases that recognize D-amino acids.

In certain embodiments the peptides are chemically synthesized by any ofa number of fluid or solid phase peptide synthesis techniques known tothose of skill in the art. Solid phase synthesis in which the C-terminalamino acid of the sequence is attached to an insoluble support followedby sequential addition of the remaining amino acids in the sequence is apreferred method for the chemical synthesis of the polypeptides of thisinvention. Techniques for solid phase synthesis are well known to thoseof skill in the art and are described, for example, by Barany andMerrifield (1963) Solid-Phase Peptide Synthesis; pp. 3-284 in ThePeptides: Analysis, Synthesis, Biology. Vol. 2: Special Methods inPeptide Synthesis, Part A.; Merrifield et al. (1963) J. Am. Chem. Soc.,85: 2149-2156, and Stewart et al. (1984) Solid Phase Peptide Synthesis,2nd ed. Pierce Chem. Co., Rockford, Ill.

In one embodiment, the peptides can be synthesized by the solid phasepeptide synthesis procedure using a benzhydrylamine resin (BeckmanBioproducts, 0.59 mmol of NH₂/g of resin) as the solid support. The COOHterminal amino acid (e.g., t-butylcarbonyl-Phe) is attached to the solidsupport through a 4-(oxymethyl)phenacetyl group. This is a more stablelinkage than the conventional benzyl ester linkage, yet the finishedpeptide can still be cleaved by hydrogenation. Transfer hydrogenationusing formic acid as the hydrogen donor can be used for this purpose.

It is noted that in the chemical synthesis of peptides, particularlypeptides comprising D amino acids, the synthesis usually produces anumber of truncated peptides in addition to the desired full-lengthproduct. Thus, the peptides are typically purified using, e.g., HPLC.

D-amino acids, beta amino acids, non-natural amino acids, and the likecan be incorporated at one or more positions in the peptide simply byusing the appropriately derivatized amino acid residue in the chemicalsynthesis. Modified residues for solid phase peptide synthesis arecommercially available from a number of suppliers (see, e.g., AdvancedChem Tech, Louisville; Nova Biochem, San Diego; Sigma, St Louis; BachemCalifornia Inc., Torrance, etc.). The D-form and/or otherwise modifiedamino acids can be completely omitted or incorporated at any position inthe peptide as desired. Thus, for example, in certain embodiments, thepeptide can comprise a single modified acid, while in other embodiments,the peptide comprises at least two, generally at least three, moregenerally at least four, most generally at least five, preferably atleast six, more preferably at least seven or even all modified aminoacids. In certain embodiments, essentially every amino acid is a D-formamino acid.

As indicated above, the peptides and/or fusion proteins of thisinvention can also be recombinantly expressed. Accordingly, in certainembodiments, the antimicrobial peptides and/or targeting moieties,and/or fusion proteins of this invention are synthesized usingrecombinant expression systems. Generally this involves creating a DNAsequence that encodes the desired peptide or fusion protein, placing theDNA in an expression cassette under the control of a particularpromoter, expressing the peptide or fusion protein in a host, isolatingthe expressed peptide or fusion protein and, if required, renaturing thepeptide or fusion protein.

DNA encoding the peptide(s) or fusion protein(s) described herein can beprepared by any suitable method as described above, including, forexample, cloning and restriction of appropriate sequences or directchemical synthesis.

This nucleic acid can be easily ligated into an appropriate vectorcontaining appropriate expression control sequences (e.g. promoter,enhancer, etc.), and, optionally, containing one or more selectablemarkers (e.g. antibiotic resistance genes).

The nucleic acid sequences encoding the peptides or fusion proteinsdescribed herein can be expressed in a variety of host cells, including,but not limited to, E. coli, other bacterial hosts, yeast, fungus, andvarious higher eukaryotic cells such as insect cells (e.g. SF3), theCOS, CHO and HeLa cells lines and myeloma cell lines. The recombinantprotein gene will typically be operably linked to appropriate expressioncontrol sequences for each host. For E. coli this can include a promotersuch as the T7, trp, or lambda promoters, a ribosome binding site andpreferably a transcription termination signal. For eukaryotic cells, thecontrol sequences can include a promoter and often an enhancer (e.g., anenhancer derived from immunoglobulin genes, SV40, cytomegalovirus,etc.), and a polyadenylation sequence, and may include splice donor andacceptor sequences.

The plasmids can be transferred into the chosen host cell by well-knownmethods such as calcium chloride transformation for E. coli and calciumphosphate treatment or electroporation for mammalian cells. Cellstransformed by the plasmids can be selected by resistance to antibioticsconferred by genes contained on the plasmids, such as the amp, gpt, neoand hyg genes.

Once expressed, the recombinant peptide(s) or fusion protein(s) can bepurified according to standard procedures of the art, including ammoniumsulfate precipitation, affinity columns, column chromatography, gelelectrophoresis and the like (see, generally, R. Scopes, (1982) ProteinPurification, Springer-Verlag, N.Y.; Deutscher (1990) Methods inEnzymology Vol. 182: Guide to Protein Purification., Academic Press,Inc. N.Y.). Substantially pure compositions of at least about 90 to 95%homogeneity are preferred, and 98 to 99% or more homogeneity are mostpreferred.

One of skill in the art would recognize that after chemical synthesis,biological expression, or purification, the peptide(s) or fusionprotein(s) may possess a conformation substantially different thandesired native conformation. In this case, it may be necessary todenature and reduce the peptide or fusion protein and then to cause themolecule to re-fold into the preferred conformation. Methods of reducingand denaturing proteins and inducing re-folding are well known to thoseof skill in the art (see, e.g., Debinski et al. (1993) J. Biol. Chem.,268: 14065-14070; Kreitman and Pastan (1993) Bioconjug. Chem., 4:581-585; and Buchner, et al., (1992) Anal. Biochem., 205: 263-270).Debinski et al., for example, describes the denaturation and reductionof inclusion body proteins in guanidine-DTE. The protein is thenrefolded in a redox buffer containing oxidized glutathione andL-arginine.

One of skill would recognize that modifications can be made to thepeptide(s) and/or fusion protein(s) proteins without diminishing theirbiological activity. Some modifications may be made to facilitate thecloning, expression, or incorporation of the targeting molecule into afusion protein. Such modifications are well known to those of skill inthe art and include, for example, a methionine added at the aminoterminus to provide an initiation site, or additional amino acids (e.g.,poly His) placed on either terminus to create conveniently locatedrestriction sites or termination codons or purification sequences.

Protecting Groups.

While the various peptides (e.g., targeting peptides, antimicrobialpeptides, STAMPs) described herein may be shown with no protectinggroups, in certain embodiments they can bear one, two, three, four, ormore protecting groups. In various embodiments, the protecting groupscan be coupled to the C- and/or N-terminus of the peptide(s) and/or toone or more internal residues comprising the peptide(s) (e.g., one ormore R-groups on the constituent amino acids can be blocked). Thus, forexample, in certain embodiments, any of the peptides described hereincan bear, e.g., an acetyl group protecting the amino terminus and/or anamide group protecting the carboxyl terminus. One example of such aprotected peptide is the 1845L6-21 STAMP having the amino acid sequenceKFINGVLSQFVLERKPYPKLFKFLRKHLL* (SEQ ID NO:1953), where the asteriskindicates an amidated carboxyl terminus. Of course, this protectinggroup can be can be eliminated and/or substituted with anotherprotecting group as described herein.

Without being bound by a particular theory, it was discovered thataddition of a protecting group, particularly to the carboxyl and incertain embodiments the amino terminus can improve the stability andefficacy of the peptide.

A wide number of protecting groups are suitable for this purpose. Suchgroups include, but are not limited to acetyl, amide, and alkyl groupswith acetyl and alkyl groups being particularly preferred for N-terminalprotection and amide groups being preferred for carboxyl terminalprotection. In certain particularly preferred embodiments, theprotecting groups include, but are not limited to alkyl chains as infatty acids, propionyl, formyl, and others. Particularly preferredcarboxyl protecting groups include amides, esters, and ether-formingprotecting groups. In one preferred embodiment, an acetyl group is usedto protect the amino terminus and an amide group is used to protect thecarboxyl terminus. These blocking groups enhance the helix-formingtendencies of the peptides. Certain particularly preferred blockinggroups include alkyl groups of various lengths, e.g., groups having theformula: CH₃—(CH₂)_(n)—CO— where n ranges from about 1 to about 20,preferably from about 1 to about 16 or 18, more preferably from about 3to about 13, and most preferably from about 3 to about 10.

In certain embodiments, the protecting groups include, but are notlimited to alkyl chains as in fatty acids, propionyl, formyl, andothers. Particularly preferred carboxyl protecting groups includeamides, esters, and ether-forming protecting groups. In one embodiment,an acetyl group is used to protect the amino terminus and/or an aminogroup is used to protect the carboxyl terminus (i.e., amidated carboxylterminus). In certain embodiments blocking groups include alkyl groupsof various lengths, e.g., groups having the formula: CH₃—(CH₂)_(n)—CO—where n ranges from about 3 to about 20, preferably from about 3 toabout 16, more preferably from about 3 to about 13, and most preferablyfrom about 3 to about 10.

In certain embodiments, the acid group on the C-terminal can be blockedwith an alcohol, aldehyde or ketone group and/or the N-terminal residuecan have the natural amide group, or be blocked with an acyl, carboxylicacid, alcohol, aldehyde, or ketone group.

Other protecting groups include, but are not limited to Fmoc,t-butoxycarbonyl (t-BOC), 9-fluoreneacetyl group, 1-fluorenecarboxylicgroup, 9-florenecarboxylic group, 9-fluorenone-1-carboxylic group,benzyloxycarbonyl, xanthyl (Xan), trityl (Trt), 4-methyltrityl (Mtt),4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr),Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl(Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl(MeBzl), 4-methoxybenzyl (MeOBl), benzyloxy (BzlO), benzyl (Bzl),benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl—Z),2-bromobenzyloxycarbonyl (2-Br—Z), Benzyloxymethyl (Bom), cyclohexyloxy(cHxO), t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), Acetyl(Ac), and Trifluoroacetyl (TFA).

Protecting/blocking groups are well known to those of skill as aremethods of coupling such groups to the appropriate residue(s) comprisingthe peptides of this invention (see, e.g., Greene et al., (1991)Protective Groups in Organic Synthesis, 2nd ed., John Wiley & Sons,Inc., Somerset, N.J.). In an illustrative embodiment, for example,acetylation is accomplished during the synthesis when the peptide is onthe resin using acetic anhydride. Amide protection can be achieved bythe selection of a proper resin for the synthesis. For example, a rinkamide resin can be used. After the completion of the synthesis, thesemipermanent protecting groups on acidic bifunctional amino acids suchas Asp and Glu and basic amino acid Lys, hydroxyl of Tyr are allsimultaneously removed. The peptides released from such a resin usingacidic treatment comes out with the n-terminal protected as acetyl andthe carboxyl protected as NH₂ and with the simultaneous removal of allof the other protecting groups.

While amino acid sequences are disclosed herein, amino acid sequencescomprising, one or more protecting groups, e.g., as described above (orany other commercially available protecting groups for amino acids used,e.g., in boc or fmoc peptide synthesis) are also contemplated.

Peptide Circularization.

In certain embodiments the peptides described herein (e.g., AMPs,compound AMPs, STAMPs, etc.) are circularized/cyclized to produce cyclicpeptides. Cyclic peptides, as contemplated herein, include head/tail,head/side chain, tail/side chain, and side chain/side chain cyclizedpeptides. In addition, peptides contemplated herein include homodet,containing only peptide bonds, and heterodet containing in additiondisulfide, ester, thioester-bonds, or other bonds.

The cyclic peptides can be prepared using virtually any art-knowntechnique for the preparation of cyclic peptides. For example, thepeptides can be prepared in linear or non-cyclized form usingconventional solution or solid phase peptide syntheses and cyclizedusing standard chemistries. Preferably, the chemistry used to cyclizethe peptide will be sufficiently mild so as to avoid substantiallydegrading the peptide. Suitable procedures for synthesizing the peptidesdescribed herein as well as suitable chemistries for cyclizing thepeptides are well known in the art.

In various embodiments cyclization can be achieved via direct couplingof the N- and C-terminus to form a peptide (or other) bond, but can alsooccur via the amino acid side chains. Furthermore it can be based on theuse of other functional groups, including but not limited to amino,hydroxy, sulfhydryl, halogen, sulfonyl, carboxy, and thiocarboxy. Thesegroups can be located at the amino acid side chains or be attached totheir N- or C-terminus.

Accordingly, it is to be understood that the chemical linkage used tocovalently cyclize the peptides of the invention need not be an amidelinkage. In many instances it may be desirable to modify the N- andC-termini of the linear or non-cyclized peptide so as to provide, forexample, reactive groups that may be cyclized under mild reactionconditions. Such linkages include, by way of example and not limitationamide, ester, thioester, CH₂—NH, etc. Techniques and reagents forsynthesizing peptides having modified termini and chemistries suitablefor cyclizing such modified peptides are well-known in the art.

Alternatively, in instances where the ends of the peptide areconformationally or otherwise constrained so as to make cyclizationdifficult, it may be desirable to attach linkers to the N- and/orC-termini to facilitate peptide cyclization. Of course, it will beappreciated that such linkers will bear reactive groups capable offorming covalent bonds with the termini of the peptide. Suitable linkersand chemistries are well-known in the art and include those previouslydescribed.

Cyclic peptides and depsipeptides (heterodetic peptides that includeester (depside) bonds as part of their backbone) have been wellcharacterized and show a wide spectrum of biological activity. Thereduction in conformational freedom brought about by cyclization oftenresults in higher receptor-binding affinities. Frequently in thesecyclic compounds, extra conformational restrictions are also built in,such as the use of D- and N-alkylated-amino acids, α,β-dehydro aminoacids or α,α-disubstituted amino acid residues.

Methods of forming disulfide linkages in peptides are well known tothose of skill in the art (see, e.g., Eichler and Houghten (1997)Protein Pept. Lett. 4: 157-164).

Reference may also be made to Marlowe (1993) Biorg. Med. Chem. Lett. 3:437-44 who describes peptide cyclization on TFA resin usingtrimethylsilyl (TMSE) ester as an orthogonal protecting group; Pallinand Tam (1995) J. Chem. Soc. Chem. Comm. 2021-2022) who describe thecyclization of unprotected peptides in aqueous solution by oximeformation; Algin et al. (1994) Tetrahedron Lett. 35: 9633-9636 whodisclose solid-phase synthesis of head-to-tail cyclic peptides vialysine side-chain anchoring; Kates et al. (1993) Tetrahedron Lett. 34:1549-1552 who describe the production of head-to-tail cyclic peptides bythree-dimensional solid phase strategy; Tumelty et al. (1994) J. Chem.Soc. Chem. Comm. 1067-1068, who describe the synthesis of cyclicpeptides from an immobilized activated intermediate, where activation ofthe immobilized peptide is carried out with N-protecting group intactand subsequent removal leading to cyclization; McMurray et al. (1994)Peptide Res. 7: 195-206) who disclose head-to-tail cyclization ofpeptides attached to insoluble supports by means of the side chains ofaspartic and glutamic acid; Hruby et al. (1994) Reactive Polymers 22:231-241) who teach an alternate method for cyclizing peptides via solidsupports; and Schmidt and Langer (1997) J. Peptide Res. 49: 67-73, whodisclose a method for synthesizing cyclotetrapeptides andcyclopentapeptides.

These methods of peptide cyclization are illustrative and non-limiting.Using the teaching provide herein, other cyclization methods will beavailable to one of skill in the art.

Joining Targeting Moieties to Effectors.

Chemical Conjugation.

Chimeric moieties are formed by joining one or more of the targetingmoieties described herein to one or more effectors. In certainembodiments the targeting moieties are attached directly to theeffector(s) via naturally occurring reactive groups or the targetingmoiety and/or the effector(s) can be functionalized to provide suchreactive groups.

In various embodiments the targeting moieties are attached toeffector(s) via one or more linking agents. Thus, in various embodimentsthe targeting moieties and the effector(s) can be conjugated via asingle linking agent or multiple linking agents. For example, thetargeting moiety and the effector can be conjugated via a singlemultifunctional (e.g., bi-, tri-, or tetra-) linking agent or a pair ofcomplementary linking agents. In another embodiment, the targetingmoiety and the effector are conjugated via two, three, or more linkingagents. Suitable linking agents include, but are not limited to, e.g.,functional groups, affinity agents, stabilizing groups, and combinationsthereof.

In certain embodiments the linking agent is or comprises a functionalgroup. Functional groups include monofunctional linkers comprising areactive group as well as multifunctional crosslinkers comprising two ormore reactive groups capable of forming a bond with two or moredifferent functional targets (e.g., labels, proteins, macromolecules,semiconductor nanocrystals, or substrate). In some preferredembodiments, the multifunctional crosslinkers are heterobifunctionalcrosslinkers comprising two or more different reactive groups.

Suitable reactive groups include, but are not limited to thiol (—SH),carboxylate (COOH), carboxyl (—COOH), carbonyl, amine (NH₂), hydroxyl(—OH), aldehyde (—CHO), alcohol (ROH), ketone (R₂CO), active hydrogen,ester, sulfhydryl (SH), phosphate (—PO₃), or photoreactive moieties.Amine reactive groups include, but are not limited to e.g.,isothiocyanates, isocyanates, acyl azides, NHS esters, sulfonylchlorides, aldehydes and glyoxals, epoxides and oxiranes, carbonates,arylating agents, imidoesters, carbodiimides, and anhydrides.Thiol-reactive groups include, but are not limited to e.g., haloacetyland alkyl halide derivates, maleimides, aziridines, acryloylderivatives, arylating agents, and thiol-disulfides exchange reagents.Carboxylate reactive groups include, but are not limited to e.g.,diazoalkanes and diazoacetyl compounds, such as carbonyldiimidazoles andcarbodiimides. Hydroxyl reactive groups include, but are not limited toe.g., epoxides and oxiranes, carbonyldiimidazole, oxidation withperiodate, N,N′-disuccinimidyl carbonate or N-hydroxylsuccimidylchloroformate, enzymatic oxidation, alkyl halogens, and isocyanates.Aldehyde and ketone reactive groups include, but are not limited toe.g., hydrazine derivatives for schiff base formation or reductionamination. Active hydrogen reactive groups include, but are not limitedto e.g., diazonium derivatives for mannich condensation and iodinationreactions. Photoreactive groups include, but are not limited to e.g.,aryl azides and halogenated aryl azides, benzophenones, diazo compounds,and diazirine derivatives.

Other suitable reactive groups and classes of reactions useful informing chimeric moieties include those that are well known in the artof bioconjugate chemistry. Currently favored classes of reactionsavailable with reactive chelates are those which proceed underrelatively mild conditions. These include, but are not limited to,nucleophilic substitutions (e.g., reactions of amines and alcohols withacyl halides, active esters), electrophilic substitutions (e.g., enaminereactions), and additions to carbon-carbon and carbon-heteroatommultiple bonds (e.g., Michael reaction, Diels-Alder addition). These andother useful reactions are discussed in, for example, March (1985)Advanced Organic Chemistry, 3rd Ed., John Wiley & Sons, New York,Hermanson (1996) Bioconjugate Techniques, Academic Press, San Diego; andFeeney et al. (1982) Modification of Proteins; Advances in ChemistrySeries, Vol. 198, American Chemical Society, Washington, D.C.

In certain embodiments, the linking agent comprises a chelator. Forexample, the chelator comprising the molecule, DOTA(DOTA=1,4,7,10-tetrakis(carboxymethyl)-1,4,7,10-tetraazacyclododecane),can readily be labeled with a radiolabel, such as Gd³⁺ and ⁶⁴Cu,resulting in Gd³⁺-DOTA and ⁶⁴Cu-DOTA respectively, attached to thetargeting moiety. Other suitable chelates are known to those of skill inthe art, for example, 1,4,7-triazacyclononane-N,N′,N″-triacetic acid(NOTA) derivatives being among the most well known (see, e.g., Lee etal. (1997) Nucl Med. Biol. 24: 2225-23019).

A “linker” or “linking agent” as used herein, is a molecule that is usedto join two or more molecules. In certain embodiments the linker istypically capable of forming covalent bonds to both molecule(s) (e.g.,the targeting moiety and the effector). Suitable linkers are well knownto those of skill in the art and include, but are not limited to,straight or branched-chain carbon linkers, heterocyclic carbon linkers,or peptide linkers. In certain embodiments the linkers can be joined tothe constituent amino acids through their side groups (e.g., through adisulfide linkage to cysteine). However, in certain embodiments, thelinkers will be joined to the alpha carbon amino and carboxyl groups ofthe terminal amino acids.

A bifunctional linker having one functional group reactive with a groupon one molecule (e.g., a targeting peptide), and another group reactiveon the other molecule (e.g., an antimicrobial peptide), can be used toform the desired conjugate. Alternatively, derivatization can beperformed to provide functional groups. Thus, for example, proceduresfor the generation of free sulfhydryl groups on peptides are also known(See U.S. Pat. No. 4,659,839).

In certain embodiments the linking agent is a heterobifunctionalcrosslinker comprising two or more different reactive groups that form aheterocyclic ring that can interact with a peptide. For example, aheterobifunctional crosslinker such as cysteine may comprise an aminereactive group and a thiol-reactive group can interact with an aldehydeon a derivatized peptide. Additional combinations of reactive groupssuitable for heterobifunctional crosslinkers include, for example,amine- and sulfhydryl reactive groups; carbonyl and sulfhydryl reactivegroups; amine and photoreactive groups; sulfhydryl and photoreactivegroups; carbonyl and photoreactive groups; carboxylate and photoreactivegroups; and arginine and photoreactive groups. In one embodiment, theheterobifunctional crosslinker is SMCC.

Many procedures and linker molecules for attachment of various moleculesto peptides or proteins are known (see, e.g., European PatentApplication No. 188,256; U.S. Pat. Nos. 4,671,958, 4,659,839, 4,414,148,4,699,784; 4,680,338; 4,569,789; and 4,589,071; and Borlinghaus et al.(1987) Cancer Res. 47: 4071-4075). Illustrative linking protocols areprovided herein in Examples 2 and 3.

Fusion Proteins.

In certain embodiments where the targeting moiety and effector are bothpeptides or both comprise peptides, the chimeric moiety can bechemically synthesized or recombinantly expressed as a fusion protein(i.e., a chimeric fusion protein).

In certain embodiments the chimeric fusion proteins are synthesizedusing recombinant DNA methodology. Generally this involves creating aDNA sequence that encodes the fusion protein, placing the DNA in anexpression cassette under the control of a particular promoter,expressing the protein in a host, isolating the expressed protein and,if required, renaturing the protein.

DNA encoding the fusion proteins can be prepared by any suitable method,including, for example, cloning and restriction of appropriate sequencesor direct chemical synthesis by methods such as the phosphotriestermethod of Narang et al. (1979) Meth. Enzymol. 68: 90-99; thephosphodiester method of Brown et al. (1979) Meth. Enzymol. 68: 109-151;the diethylphosphoramidite method of Beaucage et al. (1981) Tetra.Lett., 22: 1859-1862; and the solid support method of U.S. Pat. No.4,458,066.

Chemical synthesis produces a single stranded oligonucleotide. This canbe converted into double stranded DNA by hybridization with acomplementary sequence or by polymerization with a DNA polymerase usingthe single strand as a template. One of skill would recognize that whilechemical synthesis of DNA is limited to sequences of about 100 bases,longer sequences can be obtained by the ligation of shorter sequences.

Alternatively, subsequences can be cloned and the appropriatesubsequences cleaved using appropriate restriction enzymes. Thefragments can then be ligated to produce the desired DNA sequence.

In certain embodiments, DNA encoding fusion proteins of the presentinvention may be cloned using DNA amplification methods such aspolymerase chain reaction (PCR). Thus, for example, the nucleic acidencoding a targeting antibody, a targeting peptide, and the like is PCRamplified, using a sense primer containing the restriction site for NdeIand an antisense primer containing the restriction site for HindIII.This produces a nucleic acid encoding the targeting sequence and havingterminal restriction sites. Similarly an effector and/oreffector/linker/spacer can be provided having complementary restrictionsites. Ligation of sequences and insertion into a vector produces avector encoding the fusion protein.

While the targeting moieties and effector(s) can be directly joinedtogether, one of skill will appreciate that they can be separated by apeptide spacer/linker consisting of one or more amino acids. Generallythe spacer will have no specific biological activity other than to jointhe proteins or to preserve some minimum distance or other spatialrelationship between them. However, the constituent amino acids of thespacer may be selected to influence some property of the molecule suchas the folding, net charge, or hydrophobicity.

The nucleic acid sequences encoding the fusion proteins can be expressedin a variety of host cells, including E. coli, other bacterial hosts,yeast, and various higher eukaryotic cells such as the COS, CHO and HeLacells lines and myeloma cell lines. The recombinant protein gene will beoperably linked to appropriate expression control sequences for eachhost. For E. coli this includes a promoter such as the T7, trp, orlambda promoters, a ribosome binding site and preferably a transcriptiontermination signal. For eukaryotic cells, the control sequences willinclude a promoter and preferably an enhancer derived fromimmunoglobulin genes, SV40, cytomegalovirus, etc., and a polyadenylationsequence, and may include splice donor and acceptor sequences.

The plasmids can be transferred into the chosen host cell by well-knownmethods such as calcium chloride transformation for E. coli and calciumphosphate treatment or electroporation for mammalian cells. Cellstransformed by the plasmids can be selected by resistance to antibioticsconferred by genes contained on the plasmids, such as the amp, gpt, neoand hyg genes.

Once expressed, the recombinant fusion proteins can be purifiedaccording to standard procedures of the art, including ammonium sulfateprecipitation, affinity columns, column chromatography, gelelectrophoresis and the like (see, generally, R. Scopes (1982) ProteinPurification, Springer-Verlag, N.Y.; Deutscher (1990) Methods inEnzymology Vol. 182: Guide to Protein Purification., Academic Press,Inc. N.Y.). Substantially pure compositions of at least about 90 to 95%homogeneity are preferred, and 98 to 99% or more homogeneity are mostpreferred for pharmaceutical uses. Once purified, partially or tohomogeneity as desired, the polypeptides may then be usedtherapeutically.

One of skill in the art would recognize that after chemical synthesis,biological expression, or purification, the fusion protein may possess aconformation substantially different than the native conformations ofthe constituent polypeptides. In this case, it may be necessary todenature and reduce the polypeptide and then to cause the polypeptide tore-fold into the preferred conformation. Methods of reducing anddenaturing proteins and inducing re-folding are well known to those ofskill in the art (See, Debinski et al. (1993) J. Biol. Chem., 268:14065-14070; Kreitman and Pastan (1993) Bioconjug. Chem., 4: 581-585;and Buchner, et al. (1992) Anal. Biochem., 205: 263-270).

One of skill would recognize that modifications can be made to thefusion proteins without diminishing their biological activity. Somemodifications may be made to facilitate the cloning, expression, orincorporation of the targeting molecule into a fusion protein. Suchmodifications are well known to those of skill in the art and include,for example, a methionine added at the amino terminus to provide aninitiation site, or additional amino acids placed on either terminus tocreate conveniently located restriction sites or termination codons.

As indicated above, in various embodiments a peptide linker/spacer isused to join the one or more targeting moieties to one or moreeffector(s). In various embodiments the peptide linker is relativelyshort, typically less than about 10 amino acids, preferably less thanabout 8 amino acids and more preferably about 3 to about 5 amino acids.Suitable illustrative linkers include, but are not limited to PSGSP((SEQ ID NO:1932), ASASA (SEQ ID NO: 1933), or GGG (SEQ ID NO: 1934). Incertain embodiments longer linkers such as (GGGGS)₃ (SEQ ID NO:1935) canbe used. Illustrative peptide linkers and other linkers are shown inTable 11.

TABLE 11 Illustrative peptide and non-peptide linkers. Linker SEQ ID NO:AAA 1936 GGG 1937 SGG 1938 GGSGGS 1939 SAT 1940 PYP 1941 PSPSP 1942 ASA1943 ASASA 1944 PSPSP 1945 KKKK 1946 RRRR 1947 (Gly₄Ser)₃ 1948 GGGG 1954GGGGS 1955 GGGGS GGGGS 1956 GGGGS GGGGS GGGGS GGGGS 1957 GGGGS GGGGSGGGGS GGGGS GGGGS 1958 GGGGS GGGGS GGGGS GGGGS GGGGS GGGGS 19592-nitrobenzene or O-nitrobenzyl Nitropyridyl disulfideDioleoylphosphatidylethanolamine (DOPE) S-acetylmercaptosuccinic acid1,4,7,10-tetraazacyclododecane-1,4,7,10-tetracetic acid (DOTA)β-glucuronide and β-glucuronide variants Poly(alkylacrylic acid)Benzene-based linkers (for example: 2,5-Bis(hexyloxy)-1,4-bis[2,5-bis(hexyloxy)-4-formyl-phenylenevinylene]benzene) and like moleculesDisulfide linkages Poly(amidoamine) or like dendrimers linking multipletarget and killing peptides in one molecule Carbon nanotubes Hydrazoneand hydrazone variant linkers PEG of any chain length Succinate,formate, acetate butyrate, other like organic acids Aldols, alcohols, orenols Peroxides alkane or alkene groups of any chain length One or moreporphyrin or dye molecules containing free amide and carboxylic acidgroups One or more DNA or RNA nucleotides, including polyamine andpolycarboxyl-containing variants Inulin, sucrose, glucose, or othersingle, di or polysaccharides Linoleic acid or other polyunsaturatedfatty acids Variants of any of the above linkers containing halogen orthiol groups (all amino-acid-based linkers could be L, D, combinationsof L and D forms, β-form, PEG backbone, and the like)

Multiple Targeting Moieties and/or Effectors.

As indicated above, in certain embodiments, the chimeric moietiesdescribed herein comprise multiple targeting moieties attached to asingle effector or multiple effectors attached to a single targetingmoiety, or multiple targeting moieties attached to multiple effectors.

Where the chimeric construct is a fusion protein this is easilyaccomplished by providing multiple domains that are targeting domainsattached to one or more effector domains. FIG. 12 schematicallyillustrates a few, but not all, configurations. In various embodimentsthe multiple targeting domains and/or multiple effector domains can beattached to each other directly or can be separated by linkers (e.g.,amino acid or peptide linkers as described above).

When the chimeric construct is a chemical conjugate linear or branchedconfigurations (e.g., as illustrated in FIG. 12) are readily produced byusing branched or multifunctional linkers and/or a plurality ofdifferent linkers.

III. Administration and Formulations.

Pharmaceutical Formulations.

In certain embodiments, the antimicrobial peptides and/or the chimericconstructs (e.g., targeting moieties attached to antimicrobialpeptide(s), targeting moieties attached to detectable label(s), etc.)are administered to a mammal in need thereof, to a cell, to a tissue, toa composition (e.g., a food, etc.). In various embodiments thecompositions can be administered to detect and/or locate, and/orquantify the presence of particular microorganisms, microorganismpopulations, biofilms comprising particular microorganisms, and thelike. In various embodiments the compositions can be administered toinhibit particular microorganisms, microorganism populations, biofilmscomprising particular microorganisms, and the like.

These active agents (antimicrobial peptides and/or chimeric moieties)can be administered in the “native” form or, if desired, in the form ofsalts, esters, amides, prodrugs, derivatives, and the like, provided thesalt, ester, amide, prodrug or derivative is suitable pharmacologically,i.e., effective in the present method(s). Salts, esters, amides,prodrugs and other derivatives of the active agents can be preparedusing standard procedures known to those skilled in the art of syntheticorganic chemistry and described, for example, by March (1992) AdvancedOrganic Chemistry; Reactions, Mechanisms and Structure, 4th Ed. N.Y.Wiley-Interscience.

Methods of formulating such derivatives are known to those of skill inthe art. For example, the disulfide salts of a number of delivery agentsare described in PCT Publication WO 00/059863 which is incorporatedherein by reference. Similarly, acid salts of therapeutic peptides,peptoids, or other mimetics, and can be prepared from the free baseusing conventional methodology that typically involves reaction with asuitable acid. Generally, the base form of the drug is dissolved in apolar organic solvent such as methanol or ethanol and the acid is addedthereto. The resulting salt either precipitates or can be brought out ofsolution by addition of a less polar solvent. Suitable acids forpreparing acid addition salts include, but are not limited to bothorganic acids, e.g., acetic acid, propionic acid, glycolic acid, pyruvicacid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid,mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, and the like, as well asinorganic acids, e.g., hydrochloric acid, hydrobromic acid, sulfuricacid, nitric acid, phosphoric acid, and the like. An acid addition saltcan be reconverted to the free base by treatment with a suitable base.Certain particularly preferred acid addition salts of the active agentsherein include halide salts, such as may be prepared using hydrochloricor hydrobromic acids. Conversely, preparation of basic salts of theactive agents of this invention are prepared in a similar manner using apharmaceutically acceptable base such as sodium hydroxide, potassiumhydroxide, ammonium hydroxide, calcium hydroxide, trimethylamine, or thelike. In certain embodiments basic salts include alkali metal salts,e.g., the sodium salt, and copper salts.

For the preparation of salt forms of basic drugs, the pKa of thecounterion is preferably at least about 2 pH lower than the pKa of thedrug. Similarly, for the preparation of salt forms of acidic drugs, thepKa of the counterion is preferably at least about 2 pH higher than thepKa of the drug. This permits the counterion to bring the solution's pHto a level lower than the pHmax to reach the salt plateau, at which thesolubility of salt prevails over the solubility of free acid or base.The generalized rule of difference in pKa units of the ionizable groupin the active pharmaceutical ingredient (API) and in the acid or base ismeant to make the proton transfer energetically favorable. When the pKaof the API and counterion are not significantly different, a solidcomplex may form but may rapidly disproportionate (i.e., break down intothe individual entities of drug and counterion) in an aqueousenvironment.

Preferably, the counterion is a pharmaceutically acceptable counterion.Suitable anionic salt forms include, but are not limited to acetate,benzoate, benzylate, bitartrate, bromide, carbonate, chloride, citrate,edetate, edisylate, estolate, fumarate, gluceptate, gluconate,hydrobromide, hydrochloride, iodide, lactate, lactobionate, malate,maleate, mandelate, mesylate, methyl bromide, methyl sulfate, mucate,napsylate, nitrate, pamoate (embonate), phosphate and diphosphate,salicylate and disalicylate, stearate, succinate, sulfate, tartrate,tosylate, triethiodide, valerate, and the like, while suitable cationicsalt forms include, but are not limited to aluminum, benzathine,calcium, ethylene diamine, lysine, magnesium, meglumine, potassium,procaine, sodium, tromethamine, zinc, and the like.

In various embodiments preparation of esters typically involvesfunctionalization of hydroxyl and/or carboxyl groups that are presentwithin the molecular structure of the active agent. In certainembodiments, the esters are typically acyl-substituted derivatives offree alcohol groups, i.e., moieties that are derived from carboxylicacids of the formula RCOOH where R is alkyl, and preferably is loweralkyl. Esters can be reconverted to the free acids, if desired, by usingconventional hydrogenolysis or hydrolysis procedures.

Amides can also be prepared using techniques known to those skilled inthe art or described in the pertinent literature. For example, amidesmay be prepared from esters, using suitable amine reactants, or they maybe prepared from an anhydride or an acid chloride by reaction withammonia or a lower alkyl amine.

In various embodiments, the active agents identified herein are usefulfor parenteral, topical, oral, nasal (or otherwise inhaled), rectal, orlocal administration, such as by aerosol or transdermally, for detectionand/or quantification, and or localization, and/or prophylactic and/ortherapeutic treatment of infection (e.g., microbial infection). Thecompositions can be administered in a variety of unit dosage formsdepending upon the method of administration. Suitable unit dosage forms,include, but are not limited to powders, tablets, pills, capsules,lozenges, suppositories, patches, nasal sprays, injectables, implantablesustained-release formulations, lipid complexes, etc.

The active agents (e.g., antimicrobial peptides and/or chimericconstructs) described herein can also be combined with apharmaceutically acceptable carrier (excipient) to form apharmacological composition. Pharmaceutically acceptable carriers cancontain one or more physiologically acceptable compound(s) that act, forexample, to stabilize the composition or to increase or decrease theabsorption of the active agent(s). Physiologically acceptable compoundscan include, for example, carbohydrates, such as glucose, sucrose, ordextrans, antioxidants, such as ascorbic acid or glutathione, chelatingagents, low molecular weight proteins, protection and uptake enhancerssuch as lipids, compositions that reduce the clearance or hydrolysis ofthe active agents, or excipients or other stabilizers and/or buffers.

Pharmaceutically acceptable carriers can contain one or morephysiologically acceptable compound(s) that act, for example, tostabilize the composition or to increase or decrease the absorption ofthe active agent(s). Physiologically acceptable compounds can include,for example, carbohydrates, such as glucose, sucrose, or dextrans,antioxidants, such as ascorbic acid or glutathione, chelating agents,low molecular weight proteins, protection and uptake enhancers such aslipids, compositions that reduce the clearance or hydrolysis of theactive agents, or excipients or other stabilizers and/or buffers.

Other physiologically acceptable compounds, particularly of use in thepreparation of tablets, capsules, gel caps, and the like include, butare not limited to binders, diluent/fillers, disentegrants, lubricants,suspending agents, and the like.

In certain embodiments, to manufacture an oral dosage form (e.g., atablet), an excipient (e.g., lactose, sucrose, starch, mannitol, etc.),an optional disintegrator (e.g. calcium carbonate,carboxymethylcellulose calcium, sodium starch glycollate, crospovidoneetc.), a binder (e.g. alpha-starch, gum arabic, microcrystallinecellulose, carboxymethylcellulose, polyvinylpyrrolidone,hydroxypropylcellulose, cyclodextrin, etc.), and an optional lubricant(e.g., talc, magnesium stearate, polyethylene glycol 6000, etc.), forinstance, are added to the active component or components (e.g., active)and the resulting composition is compressed. Where necessary thecompressed product is coated, e.g., known methods for masking the tasteor for enteric dissolution or sustained release. Suitable coatingmaterials include, but are not limited to ethyl-cellulose,hydroxymethylcellulose, polyoxyethylene glycol, cellulose acetatephthalate, hydroxypropylmethylcellulose phthalate, and Eudragit (Rohm &Haas, Germany; methacrylic-acrylic copolymer).

Other physiologically acceptable compounds include wetting agents,emulsifying agents, dispersing agents or preservatives that areparticularly useful for preventing the growth or action ofmicroorganisms. Various preservatives are well known and include, forexample, phenol and ascorbic acid. One skilled in the art wouldappreciate that the choice of pharmaceutically acceptable carrier(s),including a physiologically acceptable compound depends, for example, onthe route of administration of the active agent(s) and on the particularphysio-chemical characteristics of the active agent(s).

In certain embodiments the excipients are sterile and generally free ofundesirable matter. These compositions can be sterilized byconventional, well-known sterilization techniques. For various oraldosage form excipients such as tablets and capsules sterility is notrequired. The USP/NF standard is usually sufficient.

In certain therapeutic applications, the compositions of this inventionare administered, e.g., topically administered or administered to theoral or nasal cavity, to a patient suffering from infection or at riskfor infection or prophylactically to prevent dental caries or otherpathologies of the teeth or oral mucosa characterized by microbialinfection in an amount sufficient to prevent and/or cure and/or at leastpartially prevent or arrest the disease and/or its complications. Anamount adequate to accomplish this is defined as a “therapeuticallyeffective dose.” Amounts effective for this use will depend upon theseverity of the disease and the general state of the patient's health.Single or multiple administrations of the compositions may beadministered depending on the dosage and frequency as required andtolerated by the patient. In any event, the composition should provide asufficient quantity of the active agents of the formulations of thisinvention to effectively treat (ameliorate one or more symptoms in) thepatient.

The concentration of active agent(s) can vary widely, and will beselected primarily based on activity of the active ingredient(s), bodyweight and the like in accordance with the particular mode ofadministration selected and the patient's needs. Concentrations,however, will typically be selected to provide dosages ranging fromabout 0.1 or 1 mg/kg/day to about 50 mg/kg/day and sometimes higher.Typical dosages range from about 3 mg/kg/day to about 3.5 mg/kg/day,preferably from about 3.5 mg/kg/day to about 7.2 mg/kg/day, morepreferably from about 7.2 mg/kg/day to about 11.0 mg/kg/day, and mostpreferably from about 11.0 mg/kg/day to about 15.0 mg/kg/day. In certainpreferred embodiments, dosages range from about 10 mg/kg/day to about 50mg/kg/day. In certain embodiments, dosages range from about 20 mg toabout 50 mg given orally twice daily. It will be appreciated that suchdosages may be varied to optimize a therapeutic and/or prophylacticregimen in a particular subject or group of subjects.

In certain embodiments, the active agents of this invention areadministered to the oral cavity. This is readily accomplished by the useof lozenges, aerosol sprays, mouthwash, coated swabs, and the like.

In certain embodiments, the active agent(s) of this invention areadministered topically, e.g., to the skin surface, to a topical lesionor wound, to a surgical site, and the like.

In certain embodiments the active agents of this invention areadministered systemically (e.g., orally, or as an injectable) inaccordance with standard methods well known to those of skill in theart. In other preferred embodiments, the agents, can also be deliveredthrough the skin using conventional transdermal drug delivery systems,i.e., transdermal “patches” wherein the active agent(s) are typicallycontained within a laminated structure that serves as a drug deliverydevice to be affixed to the skin. In such a structure, the drugcomposition is typically contained in a layer, or “reservoir,”underlying an upper backing layer. It will be appreciated that the term“reservoir” in this context refers to a quantity of “activeingredient(s)” that is ultimately available for delivery to the surfaceof the skin. Thus, for example, the “reservoir” may include the activeingredient(s) in an adhesive on a backing layer of the patch, or in anyof a variety of different matrix formulations known to those of skill inthe art. The patch may contain a single reservoir, or it may containmultiple reservoirs.

In one embodiment, the reservoir comprises a polymeric matrix of apharmaceutically acceptable contact adhesive material that serves toaffix the system to the skin during drug delivery. Examples of suitableskin contact adhesive materials include, but are not limited to,polyethylenes, polysiloxanes, polyisobutylenes, polyacrylates,polyurethanes, and the like. Alternatively, the drug-containingreservoir and skin contact adhesive are present as separate and distinctlayers, with the adhesive underlying the reservoir which, in this case,may be either a polymeric matrix as described above, or it may be aliquid or hydrogel reservoir, or may take some other form. The backinglayer in these laminates, which serves as the upper surface of thedevice, preferably functions as a primary structural element of the“patch” and provides the device with much of its flexibility. Thematerial selected for the backing layer is preferably substantiallyimpermeable to the active agent(s) and any other materials that arepresent.

Other formulations for topical delivery include, but are not limited to,ointments, gels, sprays, fluids, and creams. Ointments are semisolidpreparations that are typically based on petrolatum or other petroleumderivatives. Creams containing the selected active agent are typicallyviscous liquid or semisolid emulsions, often either oil-in-water orwater-in-oil. Cream bases are typically water-washable, and contain anoil phase, an emulsifier and an aqueous phase. The oil phase, alsosometimes called the “internal” phase, is generally comprised ofpetrolatum and a fatty alcohol such as cetyl or stearyl alcohol; theaqueous phase usually, although not necessarily, exceeds the oil phasein volume, and generally contains a humectant. The emulsifier in a creamformulation is generally a nonionic, anionic, cationic or amphotericsurfactant. The specific ointment or cream base to be used, as will beappreciated by those skilled in the art, is one that will provide foroptimum drug delivery. As with other carriers or vehicles, an ointmentbase should be inert, stable, nonirritating and nonsensitizing.

As indicated above, various buccal, and sublingual formulations are alsocontemplated.

In certain embodiments, one or more active agents of the presentinvention can be provided as a “concentrate”, e.g., in a storagecontainer (e.g., in a premeasured volume) ready for dilution, or in asoluble capsule ready for addition to a volume of water, alcohol,hydrogen peroxide, or other diluent.

While the invention is described with respect to use in humans, it isalso suitable for animal, e.g., veterinary use. Thus certain preferredorganisms include, but are not limited to humans, non-human primates,canines, equines, felines, porcines, ungulates, largomorphs, and thelike.

The foregoing formulations and administration methods are intended to beillustrative and not limiting. It will be appreciated that, using theteaching provided herein, other suitable formulations and modes ofadministration can be readily devised.

Home Health Care Product Formulations.

In certain embodiments, one or more of the antimicrobial peptides (AMPs)and/or chimeric moieties described herein are incorporated intohealthcare formulations, e.g., for home use. Such formulations include,but are not limited to toothpaste, mouthwash, tooth whitening strips orsolutions, contact lens storage, wetting, or cleaning solutions, dentalfloss, toothpicks, toothbrush bristles, oral sprays, oral lozenges,nasal sprays, aerosolizers for oral and/or nasal application, wounddressings (e.g., bandages), and the like.

The formulation of such health products is well known to those of skill,and the antimicrobial peptides and/or chimeric constructs are simplyadded to such formulations in an effective dose (e.g., a prophylacticdose to inhibit dental carie formation, etc.).

For example, toothpaste formulations are well known to those of skill inthe art. Typically such formulations are mixtures of abrasives andsurfactants; anticaries agents, such as fluoride; tartar controlingredients, such as tetrasodium pyrophosphate and methyl vinylether/maleic anhydride copolymer; pH buffers; humectants, to preventdry-out and increase the pleasant mouth feel; and binders, to provideconsistency and shape (see, e.g., Table 12). Binders keep the solidphase properly suspended in the liquid phase to prevent separation ofthe liquid phase out of the toothpaste. They also provide body to thedentifrice, especially after extrusion from the tube onto thetoothbrush.

TABLE 12 Typical components of toothpaste. Ingredients Wt % Humectants40-70 Water  0-50 Buffers/salts/tartar control 0.5-10  Organicthickeners (gums) 0.4-2   Inorganic thickeners  0-12 Abrasives 10-50Actives (e.g., triclosan) 0.2-1.5 Surfactants 0.5-2   Flavor andsweetener 0.8-1.5 Fluoride sources provide 1000-15000 ppm fluorine.

Table 13 lists typical ingredients used in formulations; the finalcombination will depend on factors such as ingredient compatibility andcost, local customs, and desired benefits and quality to be delivered inthe product. It will be recognized that one or more antimicrobialpeptides and/or chimeric constructs described herein can simply be addedto such formulations or used in place of one or more of the otheringredients.

TABLE 13 List of typical ingredients Inorganic Tartar Control GumsThickeners Abrasives Surfactants Humectants Ingredient Sodium SilicaHydrated Sodium Glycerine Tetrasodium carboxymethyl thickeners silicalauryl pyrophosphate cellulose sulfate Cellulose Sodium Dicalcium SodiumSorbitol Gantrez S-70 ethers aluminum phosphate N-lauryl silicatesdigydrate sarcosinate Xanthan Gum Clays Calcium Pluronics PropyleneSodium tri- carbonate glycol polyphosphate Carrageenans Sodium Xylitolbicarbonate Sodium Calcium Sodium Polyethylene alginate pyrophosphatelauryl glycol sulfoacetate Carbopols Alumina

One illustrative formulation described in U.S. Pat. No. 6,113,887comprises (1) a water-soluble bactericide selected from the groupconsisting of pyridinium compounds, quaternary ammonium compounds andbiguanide compounds in an amount of 0.001% to 5.0% by weight, based onthe total weight of the composition; (2) a cationically-modifiedhydroxyethylcellulose having an average molecular weight of 1,000,000 orhigher in the hydroxyethylcellulose portion thereof and having acationization degree of 0.05 to 0.5 mol/glucose in an amount of 0.5% to5.0% by weight, based on the total weight of the composition; (3) asurfactant selected from the group consisting of polyoxyethylenepolyoxypropylene block copolymers and alkylolamide compounds in anamount of 0.5% to 13% by weight, based on the total weight of thecomposition; and (4) a polishing agent of the non-silica type in anamount of 5% to 50% by weight, based on the total weight of thecomposition. In certain embodiments, the antimicrobial peptide(s) and/orchimeric construct(s) described herein can be used in place of thebactericide or in combination with the bactericide.

Similarly, mouthwash formulations are also well known to those of skillin the art. Thus, for example, mouthwashes containing sodium fluorideare disclosed in U.S. Pat. Nos. 2,913,373, 3,975,514, and 4,548,809, andin US Patent Publications US 2003/0124068 A1, US 2007/0154410 A1, andthe like. Mouthwashes containing various alkali metal compounds are alsoknown: sodium benzoate (WO 9409752); alkali metal hypohalite (US20020114851A1); chlorine dioxide (CN 1222345); alkali metal phosphate(US 2001/0002252 A1, US 2003/0007937 A1); hydrogen sulfate/carbonate (JP8113519); cetylpyridium chloride (CPC) (see, e.g., U.S. Pat. No.6,117,417, U.S. Pat. No. 5,948,390, and JP 2004051511). Mouthwashescontaining higher alcohol (see, e.g., US 2002/0064505 A1, US2003/0175216 A1); hydrogen peroxide (see, e.g., CN 1385145); CO₂ gasbubbles (see, e.g., JP 1275521 and JP 2157215) are also known. Incertain embodiments, these and other mouthwash formulations can furthercomprise one or more of the AMPs or compound AMPs of this invention.

Contact lens storage, wetting, or cleaning solutions, dental floss,toothpicks, toothbrush bristles, oral sprays, oral lozenges, nasalsprays, and aerosolizers for oral and/or nasal application, and the likeare also well known to those of skill in the art and can readily beadapted to incorporate one or more antimicrobial peptide(s) and/orchimeric construct(s) described herein.

The foregoing home healthcare formulations and/or devices are meant tobe illustrative and not limiting. Using teaching provided herein, theantimicrobial peptide(s) and/or chimeric construct(s) described hereincan readily be incorporated into other products.

IV. Kits.

In another embodiment this invention provides kits for the inhibition ofan infection and/or for the treatment and/or prevention of dental cariesin a mammal. The kits typically comprise a container containing one ormore of the active agents (i.e., the antimicrobial peptide(s) and/orchimeric construct(s)) described herein. In certain embodiments theactive agent(s) can be provided in a unit dosage formulation (e.g.,suppository, tablet, caplet, patch, etc.) and/or may be optionallycombined with one or more pharmaceutically acceptable excipients.

In certain embodiments the kits comprise one or more of the homehealthcare product formulations described herein (e.g., toothpaste,mouthwash, tooth whitening strips or solutions, contact lens storage,wetting, or cleaning solutions, dental floss, toothpicks, toothbrushbristles, oral sprays, oral lozenges, nasal sprays, aerosolizers fororal and/or nasal application, and the like).

In certain embodiments kits are provided for detecting and/or locatingand/or quantifying certain target microorganisms and/or cells or tissuescomprising certain target microorganisms, and/or prosthesis bearingcertain target microorganisms, and/or biofilms comprising certain targetmicroorganisms. In various embodiments these kits typically comprise achimeric moiety comprising a targeting moiety and a detectable label asdescribed herein and/or a targeting moiety attached to an affinity tagfor use in a pretargeting strategy as described herein.

In addition, the kits optionally include labeling and/or instructionalmaterials providing directions (i.e., protocols) for the practice of themethods or use of the “therapeutics” or “prophylactics” or detectionreagents of this invention. Certain instructional materials describe theuse of one or more active agent(s) of this invention to therapeuticallyor prophylactically to inhibit or prevent infection and/or to inhibitthe formation of dental caries. The instructional materials may also,optionally, teach preferred dosages/therapeutic regiment, counterindications and the like.

While the instructional materials typically comprise written or printedmaterials they are not limited to such. Any medium capable of storingsuch instructions and communicating them to an end user is contemplatedby this invention. Such media include, but are not limited to electronicstorage media (e.g., magnetic discs, tapes, cartridges, chips), opticalmedia (e.g., CD ROM), and the like. Such media may include addresses tointernet sites that provide such instructional materials.

EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed invention.

Example 1 Design and Activity of a “Dual-Targeted” Antimicrobial Peptide

Numerous reports have indicated the important role of human normal florain the prevention of microbial pathogenesis and disease. Evidencesuggests that infections at mucosal surfaces result from the outgrowthof subpopulations or clusters within a microbial community, and are notlinked to one pathogenic organism alone. In order to preserve theprotective normal flora while treating the majority of infectivebacteria in the community, a tunable therapeutic is necessary that candiscriminate between benign bystanders and multiple pathogenicorganisms. Here we describe the proof-of-principle for such amulti-targeted antimicrobial: a multiple-headed specifically-targetedantimicrobial peptide (MH-STAMP). The completed MH-STAMP, M8(KH)-20,displays specific activity against targeted organisms in vitro(Pseudomonas aeruginosa and Streptococcus mutans) and can remove bothspecies from a mixed planktonic culture with little impact againstuntargeted bacteria. These results demonstrate that a functional,dual-targeted molecule can be constructed from wide-spectrumantimicrobial peptide precursor.

Introduction

For nearly 30 years antimicrobial peptides (AMPs) have been rigorouslyinvestigated as alternatives to small molecule antibiotics and potentialsolutions to the growing crisis of antibiotic resistant bacterialinfections [1, 2]. Numerous reports have characterized potential AMPsfrom natural sources, and a great body of work has been carried outdesigning “tailor-made” AMPs due to the approachable nature ofsolid-phase peptide synthesis (SPPS) [3, 4]. Several examples of thelatter have shown remarkable activities in vitro against fungi,Gram-positive and Gram-negative bacteria, as well as some envelopedviruses [5].

Unlike small molecule antibiotics that may lose activity when theirbasic structures are modified even incrementally, peptides are aconvenient canvas for molecular alteration. AMPs can be optimizedthrough the incorporation of more or less hydrophobic or charged aminoacids, which has been shown to affect selectivity for Gram-positive,Gram-negative or fungal membranes [6, 7]. Additionally, lysine residuescan be utilized to improve AMP activity per μM. In this approach,multiple AMP chains can be attached to a single peptide scaffold throughbranching from lysine epsilon-amines [8, 9]. AMP activity can bespecifically tuned through the attachment of a targeting peptide region,as described for a novel class of molecules, the specifically-targetedantimicrobial peptides, or STAMPs [10, 11]. These chimeric molecules canconsist of functionally independent targeting and killing moietieswithin a linear peptide sequence. A pathogenic bacterium recognized(i.e. bound) by the targeting peptide can be eliminated from amulti-species community with little impact to bystander normal flora. Asan extension of this concept, we hypothesized that a STAMP could beconstructed with multiple targeting peptide “heads” attached to a singleAMP by utilizing a central lysine residue branch point. Potentially,targeting “heads” could be specific for the same pathogen, or havedifferent binding profiles. Utilizing the former approach, microbialresistance evolution linked to a targeting peptide could be inhibited orreduced, as no single microbial population would have the geneticdiversity necessary to mutate multiple discrete targeting peptidereceptors in one cell [12].

Multi-headed STAMP (MH-STAMP) molecules with differing bacterial targetsmay have appeal in treating poly-microbial infections, or where it maybe advantageous to remove a cluster of biofilm constituents withoututilizing several distinct molecules; for example in the simultaneouslytreatment of dental caries and periodontitis, or in the eradication ofthe Propionibacteria spp. and Staphylococcus spp. involved in acne andskin infections, respectively.

In this example, we present the proof-of-principle design, synthesis andin vitro activity of such a MH-STAMP, M8(KH)-20. Previously, weidentified two functional STAMP targeting domains, one with specificrecognition of the cariogenic pathogen S. mutans [10], and the otherwith Pseudomonas spp.-level selectivity [13]. Conjoined to a normallywide-spectrum linear AMP, we observed antimicrobial effects directedspecifically to P. aeruginosa and S. mutans in vitro. Additionally,treatment of mixed bacterial communities with the multi-headed MH-STAMPresulted in the specific eradication of the target organisms with littleimpact on bystander population levels.

Materials and Methods

Bacterial Strains and Growth Conditions

P. aeruginosa ATCC 15692, Klebsiella pneumoniae KAY 2026 [14],Escherichia coli DH5α (pFW5, spectinomycin resistance) [15],Staphylococcus aureus Newmann [16], and Staphylococcus epidermidis ATCC35984 were cultivated under aerobic conditions at 37° C. with vigorousshaking Aerobic Gram-negative organisms were grown in Lauri-Bertaini(LB) broth and Gram-positive bacteria in Brain-heart infusion (BHI)broth. Streptococcus mutans JM11 (spectinomycin resistant, UA140background) was grown in Todd-Hewitt (TH) broth under anaerobicconditions (80% N2, 15% CO₂, 5% H2) at 37° C. [17]. All bacteria weregrown overnight to an OD600 of 0.8-1.0 prior to appropriate dilution andantimicrobial testing.

Synthesis of Multi-Head STAMP Peptides

Conventional solid-phase peptide synthesis (SPPS) methodologies wereutilized for the construction of all peptides shown in FIG. 13 (SymphonySynthesizer, PTI, Tucson, Ariz.). Chemicals, amino acids, and synthesisresins were purchased from Anaspec (San Jose, Calif.). BD2.20(FIRKFLKKWLL (SEQ ID NO:1949), amidated c-terminus, mw 1491.92), anantimicrobial peptide developed in our laboratory with robustantimicrobial activity against a number of bacterial species (Table 14),served as the root sequence to which differing targeting peptides wereattached: Firstly, BD2.20 was synthesized by SPPS (Rink-Amide-MBHAresin, 0.015 mmol), followed by the stepwise coupling of afunctionalized alkane (NH₂(CH₂)₇COOH), and an Fmoc-protected Lys(side-chain protected with 4-methyltrityl (Mtt)) to the N-terminus.Standard SPPS methods were then employed for the step-wise addition ofthe S. mutans targeting peptide M8 plus a tri-Gly linker region(TFFRFLNR-GGG (SEQ ID NO:1950)) to the N-terminal of the central Lys.After assembly of Fmoc-M8-GGG-K(Mtt)-(CH₂)₇CO-BD2.20 (SEQ ID NO:1951),the Fmoc group was removed with 25% piperidine in DMF and the N-terminalwas re-protected with an acetyl group with Ac₂O/DIEA (1:1, 20 molarexcess) for 2 hours. The Mtt-protected amino group of the central Lyswas then selectively exposed with 2% TFA in DCM (1.5 mL) for 15 minutes(three cycles of 5 min). The resulting product was reloaded into thesynthesizer and the peptide sequence built from the Lys side-chain wascompleted with standard Fmoc SPPS methods. As shown in FIG. 13, thecompleted MH-STAMP M8(KH)-20 contained the side-chain peptide KH(Pseudomonas spp.-targeting, KKHRKHRKHRKH-GGG (SEQ ID NO:1952)), whilein MH-STAMP M8(BL)-20 a peptide with no bacterial binding (data notshown), BL-1 (DAANEA-GGG), was utilized. BL(KH)-20 was constructedidentically to M8(KH)-20, utilizing BL-1 in place of M8 (FIG. 13).

TABLE 14 MICs of MH-STAMPs and component peptides. MIC (μM) P.aeruginosa E. coli K. pneumoniae S. mutans S. epidermidis S. aureusBD2.20 14.4 ± 4.40 5.47 ± 1.41 2.98 ± 0.47 2.86 ± 0.60 5.11 ± 1.58 5.625± 1.29 M8(KH)-20 11.95 ± 3.32  2.72 ± 0.59 3.13 6.25 3.13  5.64 ± 1.07M8(BL)-20 50 5.97 ± 0.94 6.88 ± 1.98 6.25 6.25 18.05 ± 6.58 BL(KH)-2027.5 ± 7.90 6.25 6.25 6.25 6.25 6.25 Average MIC with standarddeviation, n = 10 assays.

Synthesis progression was monitored by the ninhydrin test, and completedpeptides cleaved from the resin with 95% TFA utilizing appropriatescavengers, and precipitated in methyl tert-butyl ether. Purificationand MH-STAMP quality was confirmed by HPLC (Waters, Milford, Mass.)using a linear gradient of increasing mobile phase (acetonitrile 10 to90% in water with 0.1% TFA) and a Waters XBridge BEH 130 C18 column(4.6×100 mm, particle size 5 μm). Absorbance at 215 nm was utilized asthe monitoring wavelength, though 260 and 280 nm were also collected. LCspectra were analyzed with MassLynx Software v.4.1 (Waters).Matrix-assisted laser desorption ionization (MALDI) mass spectroscopywas utilized to confirm correct peptide mass (Voyager System 4291,Applied Biosystems) [18].

MIC Assay

Peptides were evaluated for basic antimicrobial activity by brothmicrodilution, as described previously [10, 11]. Briefly, ˜1×10⁵ cfu/mLbacteria were diluted in TH (S. mutans), or Mueller-Hinton (MH) broth(all other organisms) and distributed to 96-well plates.Serially-diluted (2-fold) peptides were then added and the platesincubated at 37° C. for 18-24 h. Peptide MIC was determined as theconcentration of peptide that completely inhibited organism growth whenexamined by eye (clear well). All experiments were conducted 10 times.

Post-Antibiotic Effect Assay

The activity and selectivity of MH-STAMPs after a 10 min incubation wasdetermined by growth retardation experiments against targeted anduntargeted bacteria in monocultures, as described previously [10, 11].Cells from overnight cultures were diluted to ˜5×10⁶ cfu/mL in MH (or THwith 1% sucrose for S. mutans), normalized by OD600 0.05-0.1 and seededto 96-well plates. Cultures were then grown under the appropriateconditions for 2 h (3 h for S. mutans) prior to the addition of peptidesfor 10 min. Plates were then centrifuged at 3000×g for 5 min, thesupernatants discarded, fresh medium returned (MH or TH without sucrosefor S. mutans), and incubation resumed. Bacterial growth after treatmentwas then monitored over time by OD600.

Microbial Population Shift Assay

Mixed planktonic populations of P. aeruginosa, E. coli, S. epidermidis,and S. mutans were utilized to examine the potential of MH-STAMPs todirect species composition within a culture after treatment. Sampleswere prepared containing: ˜6×10⁴ cfu/mL S. mutans, ˜2×10⁴ cfu/mL E.coli, ˜2×10⁴ cfu/mL S. epidermidis, and ˜0.5×10⁴ cfu/mL P. aeruginosa inBHI (mixed immediately before peptide addition). Peptide (10 μM) ormock-treatment (1×PBS) was then added and samples were incubated at 37°C. for 24 h under anaerobic conditions (80% N2, 15% CO₂, 5% H2). Afterincubation, samples were serially diluted (1:10) in 1×PBS and aliquotsfrom each dilution were then spotted to agar plates selective for eachspecies in the mixture: TH plus 800 μg/mL spectinomycin (S. mutans), LBplus 25 μg/mL ampicillin (P. aeruginosa), LB plus 200 μg/mLspectinomycin (E. coli), and mannitol salt agar (MSA, S. epidermidis) inorder to quantitate survivors from each species. Plates were thenincubated 37° C. under aerobic conditions (TH plates were incubatedanaerobically) and colonies counted after 24 h to determine survivors.Expected colony morphologies were observed for each species when platedon selective media. Gram stains and direct microscopic observation (fromselect isolated colonies) were undertaken to confirm species identity(data not shown). The detection limit of the assay was 200 cfu/mL.

Results

Design and Synthesis of Multi-Headed STAMPs

We constructed a prototype MH-STAMP from the well-established targetingpeptides KH (specific to Pseudomonas spp) and M8 (specific forStreptococcus mutans). The wide-spectrum antimicrobial peptide BD2.20was utilized as the base AMP for all MH171 STAMP construction. BD2.20 isa novel synthetic AMP with a cationic and amphipathic residuearrangement, which has robust MICs against a variety of Gram-negativeand Gram-positive organisms (Table 14). For the synthesis of MH-STAMPM8(KH)-20 (construct presented in FIG. 13), BD2.20 and a Lys(Mtt-protected side-chain) residue were joined via an activated alkanespacer, followed by addition of the M8 targeting peptide to theN-terminus of the product. Selective deprotection of the centralLys(Mtt) side chain was then undertaken and the KH targeting peptideattached. The correct molecular mass (4888.79) and ˜90% purity wasconfirmed by HPLC and MALDI mass spectrometry (FIG. 14).

The non-binding “blank” targeting peptide BL-1 was incorporated into thesynthesis scheme in place of KH or M8 to construct variant MH-STAMPspossessing a single functional targeting head: M8(BL)-20 and BL(KH)-20.The correct MW and acceptable purity were observed for these MH-STAMPs(FIG. 13, data not shown).

General Antimicrobial Activity of Multi-Head Constructs

After synthesis, the completed MH-STAMPs were evaluated for generalantimicrobial activity by MIC against a panel of bacteria. As shown inTable 14, the MH-STAMP constructs M8(KH)-20, BL(KH)-20, and M8(BL)-20were found to have similar activity profiles to that of BD2.20 for theorganisms examined (less than two titration steps in 10-folddifference). Additionally, we observed a difference in generalsusceptibility between P. aeruginosa and the other organisms tested,suggesting this bacterium is more resistant to BD2.20. Overall, thesedata indicate that the addition of the targeting domains to the basesequence was tolerated and did not completely inhibit the activity ofthe antimicrobial peptide.

Peptide selectivity could not be determined utilizing these methods, asSTAMPs and their parent AMP molecules often display similar MICs, buthave radically different antimicrobial kinetics and selectivity due toincreased specific-killing mediated by the targeting regions [10, 11].Therefore, we performed different experiments to test for antimicrobialselectivity and functional MH-STAMP construction.

3.3 Selectivity and Post-Antibiotic Effect of MH-STAMP Constructs

MH-STAMP antimicrobial kinetics was ascertained utilizing a variation ofthe classical post-antibiotic effect assay, which measures the abilityof an agent to affect an organism's growth after a short exposureperiod. Monocultures of MH-STAMP-targeted and untargeted organisms wereexposed to M8(KH)-20, M8(BL)-20, BL(KH)-20, or unmodified BD2.20, thenallowed to recover. As shown in FIG. 15A, S. mutans growth waseffectively retarded by M8-containing constructs (M8(KH)-20, M8(BL)-20),but was not altered by a MH-STAMP construct lacking this region(BL(KH)-20). Similarly, the growth of the other targeted bacterium, P.aeruginosa, was inhibited in a KH-dependant manner (FIG. 15B). Incomparison, the non-targeted bacteria E. coli, S. aureus, and S.epidermidis were not inhibited by treatment with any MH-STAMP and wereonly inhibited by the base antimicrobial peptide BD2.20, which displayedrobust antimicrobial activity against all examined strains. Theseresults indicate that MH-STAMPs containing KH or M8 targeting domainshave activity against P. aeruginosa or S. mutans, respectively, and notother bacteria. Furthermore, replacement of the targeting region with anon-binding peptide abolishes specific activity.

Ability of MH-STAMPs to Direct a “Population Shift” within a MixedSpecies Population

We hypothesized that potential MH-STAMP dual-functionality could affecta particular set of bacteria within a mixed population, therebypromoting the outgrowth of non-targeted organisms and “shifting” theconstituent makeup. To examine this possibility, defined mixedpopulations of planktonic cells were treated continuously and themake-up of the community examined after 24 h. As shown in FIG. 16,treatment with the wide spectrum AMP BD2.20 resulted in a significantloss of recoverable cfu/mL after 24 h from all species in the mixture.Treatment with M8(KH)-20 was found to alter this pattern; we observed˜1×10⁵ cfu/mL surviving E. coli and S. epidermidis, but did not recoverS. mutans or P. aeruginosa cfu/mL. In BL(KH)-20 treated samples, P.aeruginosa cfu/mL were not observed, though we recovered higher thaninput cfu/mL from S. mutans and unchanged numbers of S. epidermidis andE. coli. In samples exposed to M8(BL)-20, S. mutans recoverable cfu/mLwere greatly reduced compared to input cfu/mL, while other species werenot affected or affected to a lesser extent. Interestingly, theseresults suggest that M8(KH)-20, M8(BL)-20, and BL(KH)-20 retain theirability to affect organisms recognized by the targeting regions present,even within a mixed population of bacteria.

Discussion

Our results indicate that we have successfully constructed a STAMP withdual antimicrobial specificities controlled by the targeting peptidespresent in the molecule; KH for Pseudomonas spp, M8 for S. mutans. In aclosed multi-species system (FIG. 16), the dual specificity of M8(KH)-20was readily discernable: the population of the culture “shifted” awayfrom targeted organisms after MH-STAMP treatment. The targeted bacteriawere eliminated and the population of untargeted organisms increased, tovarying degrees, above-input cfu/mL. Additionally, interruption of KH orM8 in the MH-STAMP construct with the non-binding peptide BL-1 resultedin the expected elimination of only one targeted species. These resultssupport the hypothesis that functional MH-STAMPs could be constructedfrom a wide-spectrum AMP base.

The emergence of metagenomics and the development of more sensitivemolecular diagnostics has driven an increase in the understanding ofhuman-associated microbial ecologies and host-microbe interactions[19-21]. At mucosal surfaces, it has become clear that our bodies harboran abundance of residential flora which may impact innate and humoralimmunity, nutrient availability, protection against pathogens, and evenhost physiology [22-25]. Furthermore, findings have indicated thatshifts in the diversity of normal flora are associated with negativeclinical consequences; for example the overgrowth of S. mutans in theoral cavity during cariogenesis (linked to the uptake of sucrose) or theantibiotic-assisted colonization of the intestine by Clostridiumdifficle [26, 27]. Other population shifts may be linked to axilla odor(Corynebacteria spp) [28, 29], or even host obesity. Given the quantityand diversity of microbes present, pathogenesis at mucosal surfaces isnot likely to be associated with the overgrowth of a single strain orspecies. More often, it is a population shift resulting in thepredominance of two or more species; for example the persistence ofBurkholderia cepacia and P. aeruginosa in cystic fibrosis airway orTreponema denticola and Porphymonas gingivalis and other “red cluster”organisms in gingivitis [30, 31]. In many cases (such as the latter)these species may have only distant phylogenetic relationships anddisplay differential susceptibilities to antibiotic therapies resultingin persistent disease progression despite treatment [32, 33]. Currently,available treatments for infections of mucosal surfaces are largelynon-specific (traditional small-molecule antibiotics, mechanicalremoval), and thus are not effective in retaining flora or shifting theconstituent balance back to a health-associated composition [34]. Thereis a need for a therapeutic treatment that can selectively targetmultiple pathogens, regardless of their phylogenetic relationship, andMH-STAMPs can help achieve this goal.

In monoculture experiments (FIG. 15), our results suggest that M8 or KHinclusion in the MH-STAMP drove activity towards S. mutans or P.aeruginosa, but also that the presence of a targeting domain reduced theactivity of the parent AMP BD2.20 against untargeted organisms. Incontrast, the results of our MIC assays (Table 14) indicate littledifference in activity between BD2.20 and any MH-STAMP. Againstuntargeted organisms, the M8 and KH regions are likely to have anegative, but not completely inhibitory, impact on BD2.20 activity.Given the long duration of activity and the lower inoculum size in theMIC assay (compared with experiments in FIG. 15), it is likely that allBD2.20-containing peptides could reach equal levels of growthinhibition, despite large and target-specific differences inantimicrobial speed. This pattern of results was also observed whencomparing MICs of targeted and untargeted organisms utilizing STAMPsagainst S. mutans and Pseudomonas mendocina [10, 11].

Although more rigorous studies and a more medically relevant combinationof pathogen targets is desirable, these findings indicate that it ispossible to design an antimicrobial peptide-based therapeutic withmultiple and defined fidelities in vitro. MH-STAMPs may help improvehuman health through the promotion of healthy microbial constituencies.

REFERENCES

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Enhancement of Antimicrobial Activity against Pseudomonas    aeruginosa by Coadministration of G10KHc and Tobramycin Antimicrob    Agents Chemother 2006; 50:3833-8.-   14. Sprenger G A, Lengeler J W. L-Sorbose metabolism in Klebsiella    pneumoniae and Sor+ derivatives of Escherichia coli K-12 and    chemotaxis toward sorbose. J Bacteriol 1984; 157:39-45.-   15. Podbielski A, Spellerberg B, Woischnik M, Pohl B, Lütticken R.    Novel series of plasmid vectors for gene inactivation and expression    analysis in group A streptococci (GAS). Gene 1996; 177:137-47.-   16. Duthie E S, Lorenz L L. Staphylococcal coagulase; mode of action    and antigenicity. J Gen Microbiol 1952; 6:95-107.-   17. Merritt J, Kreth J, Qi F, Sullivan R, Shi W. Non-disruptive,    real-time analyses of the metabolic status and viability of    Streptococcus mutans cells in response to antimicrobial treatments.    J Microbiol Methods 2005; 61:161-70.-   18. Anderson R C, Rehders M, Yu P L. Antimicrobial fragments of the    pro-region of cathelicidins and other immune peptides. Biotechnol    Lett 2008; 30:813-8.-   19. Aas J A, Paster B J, Stokes L N, Olsen I, Dewhirst F E. Defining    the normal bacterial flora of the oral cavity. J Clin Microbiol    2005; 43:5721-32.-   20. Boman H G. Innate immunity and the normal microflora. Immunol    Rev 2000; 173:5-16.-   21. Kreth J, Merritt J, Shi W, Qi F. Competition and coexistence    between Streptococcus mutans and Streptococcus sanguinis in the    dental biofilm. J Bacteriol 2005; 187:7193-203.-   22. Metges C C. Contribution of Microbial Amino Acids to Amino Acid    Homeostasis of the Host. J Nutr 2000; 130:1857 S-64.-   23. Sears C L. A dynamic partnership: Celebrating our gut flora.    Anaerobe 2005; 11:247-251.-   24. Lievin-Le Moal V, Servin A L. The Front Line of Enteric Host    Defense against Unwelcome Intrusion of Harmful Microorganisms:    Mucins, Antimicrobial Peptides, and Microbiota Clin Microbiol Rev    2006; 19:315-37.-   25. DiBaise J K, Zhang H, Crowell M D, Krajmalnik-Brown R, Deckert G    A, Rittmann B E. Gut microbiota and its possible relationship with    obesity. Mayo Clinic Proceedings 2008; 83:460-9.-   26. Loesche W J. Role of Streptococcus mutans in human dental decay.    Microbiol. Rev 1986; 50:353-80.-   27. Gould C V, McDonald LC. Bench-to-bedside review: Clostridium    difficile colitis. Crit. Care 2008; 12:203.-   28. Leyden J J, McGinley K J, Holzle E, Labows J N, Kligman A M. The    microbiology of the human axilla and its relationship to axillary    odor. J Invest Dermatol 1981; 77:413-6.-   29. Elsner P. Antimicrobials and the skin physiological and    pathological flora. Curr Probl Dermatol 2006; 33:35-41.-   30. Govan J R, Deretic V. 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Example 2 Synthesis of Peptide-Porphyrin Conjugate

The mixture of coupling reagent HATU (5 eq. excess, 10 mg) andpurpurin-18 (MW 564, 5 eq excess, 15 mg) in 600 mL dry dichloromethane(DCM):DMF:dimethylsulphoxide (DMSO) (1:1:1 (v/v)) was added to thepeptide resin (1 molar equivalent, 15 mg) which was swelled by placingin minimal DMF for 30 min prior to reaction. 26 μL (10 molarequivalents) DIPEA was then added to the reaction flask to initiate thereaction. The reaction mixture was protected with argon and stirred atroom temperature for 3 h.

After finishing, the reaction mixture was then passed down a sinteredglass filtered vial and extensively washed with DMF and DCM to removeall waste reagents. The resin was then dried overnight in vacuum, andcleaved with 1 ml of trifluoroacetic acid (TFA)/thioanisole/water/EDT(10/0.5/0.5/025) for 2 hr at room temperature, and the cleavage solutionwas precipitated with 10 mL methyl-tert butyl ether. The precipitate waswashed twice with the same amount of ether.

Example 3 Synthesis of Peptide-CSA Conjugate

To the fully protected peptide (solution of B43-GGG (FIDSFIRSF-GGG,0.025 mmol) and tri-Boc-CSA-15 (0.0125 mol) in 300 μL DMF, DCC (7.7 mg),HOBt (5.1 mg) and 13 μL DIEA were added in iced-bath. After stirred atroom temperature for four days, the reaction mixture was poured into 5ml water and extracted with chloroform (5×3 mL). The CHCl₃ extract wasevaporated under vacuum and dried in a lyophilizer overnight. The driedCHCl₃ extracts was then dissolved in 1 mL DCM followed by added 1 mL ofTFA in iced-bath. The reaction mixture was further stirred at roomtemperature for 2 hours and precipitated with methyl tert-butyl ether(10 mL). The precipitate was further washed once with the same amountether and dried in vacuum.

Example 4 Systemically Designed STAMPS Against S. Mutans

We previously reported a novel strategy of “targeted-killing” with thedesign of narrow-spectrum molecules known as specifically-targetedantimicrobial peptides (STAMPs). Construction of these moleculesrequires the identification and the subsequent utilization of twoconjoined yet functionally independent peptide components: the targetingand killing regions. In this study, we sought to design and synthesize alarge number of STAMPs targeting Streptococcus mutans, the primaryetiological agent of human dental caries, in order to identify candidatepeptides with increased killing speed and selectivity when compared withtheir unmodified antimicrobial peptides (AMP) precursors. Wehypothesized that a combinatorial approach utilizing a set number ofAMP, targeting, and linker regions, would be an effective method for theidentification of STAMPs with the desired level of activity. STAMPscomposed of the S. mutans binding peptide 2_(—)1 and the AMP PL-135displayed selectivity at minimal inhibitory concentrations afterincubation for 18-24 h. STAMPs where PL-135 was replaced by the B-33killing domain exhibited both selectivity and rapid killing within oneminute of exposure. These results suggest that potent and selectiveSTAMP molecules can be designed and improved via a tunable“building-block” approach.

Introduction

Pathogenic microorganisms have been a continuous source of humansuffering and mortality throughout the course of human history and havespurred the clinical development of novel therapeutics. Even today, theoverall burden of infectious disease remains high, constituting aleading (and rising) cause of death worldwide (12, 13). The conventionalmedical response to bacterial infections, small molecule antibiotics,have become less effective against emerging pathogens due to theevolution of drug-resistance stemming in part from the misuse ofantibiotics (11). To counter the rapid progression of antibioticresistance there is an urgent need for the development of novel leadcompounds for clinical applications.

Our strategy for creating new antibacterial agents is based on theaddition of a targeting peptide to an existing broad-spectrumantimicrobial peptide, thereby generating a specifically-targetedantimicrobial peptide (STAMP) selective for a particular bacterialspecies or strain. A completed STAMP consists of conjoined butfunctionally independent targeting and killing regions, separated by asmall flexible linker, all within a linear peptide sequence. The STAMPtargeting region drives enhancement of antimicrobial activity byincreasing binding to the surface of a targeted pathogen, utilizingspecific determinants such as overall membrane hydrophobicity andcharge, pheromone receptors, etc., which in turn leads to increasedselective accumulation of the killing moiety (6, 7).

As both the killing and targeting regions of the STAMP are linearpeptides, we approached the design process using a tunable combinatorialmethodology where, for example, the targeting peptide component is heldconstant, while a number of killing peptides are conjoined utilizing avariety of linker molecules, or vise versa, in order to generate alibrary of related STAMPs. Previously, we successfully demonstrated apilot version of this approach when constructing G10KHc (6), a STAMPwith Pseudomonas-spp selective activity, and when designing C16G2 (7), aSTAMP specific for Streptococcus mutans, the leading causative agent ofhuman tooth decay. In this study, synthetic targeting and antimicrobialpeptide libraries were utilized as building blocks to generate a numberof novel STAMPs with high S. mutans-selective activity. STAMPs designedby these methods were then improved through tuning the linker andkilling peptides present to yield completed lead STAMP molecules thatdemonstrated activity against S. mutans biofilms.

Materials and Methods

Reagents. Wang resin, Rink-MBHA resin, p-Benzyloxybenzyl alcohol resin(100-200 mesh), 9-fluorenylmethoxycarbonyl (Fmoc) amino acids,N-hydroxybenzotriazole hydrate (HOBT) and2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate(HBTU) were obtained from Anaspec (San Jose, Calif.). All other solventsand reagents were purchased from Fisher Scientific (Pittsburgh, Pa.) atHPLC or peptide synthesis grade.

Bacterial Growth All S. mutans (UA159) (1), Streptococcus gordoniiChallis (DL 1), Streptococcus sobrinus ATCC 33478, and Streptococcussanguinis NY101 strains were grown in Brain Heart Infusion (BHI) mediumat 37° C. under anaerobic conditions (80% N₂, 10% CO₂, 10% H₂) (6).Pseudomonas aeruginosa (PAK) (17), and Escherichia coli W3110 (20), werecultured in Luria-Bertani (LB) medium under an aerobic atmosphere at 37°C. Methicillin-resistant Staphylococcus aureus (MRSA), andvancomycin-resistant Enterococcus faecium (VRE) were grown in BHI underaerobic conditions at 37° C. (4).

Peptide Syntheses and Purification. Peptides were synthesized usingstandard solid phase (Fmoc) chemistry with an Apex 396 peptidesynthesizer (AAPPTec, Louisville, Ky.) at a 0.01 mM scale. N-terminaldeblocking during linear peptide synthesis was conducted with 0.6 mL of25% (v/v) piperidine in dimethylformamide (DMF), followed by agitationfor 27 min and wash cycles of dichlormethane DCM (1×1 mL) and NMP (7×0.8mL). Subsequent amino acid coupling cycles were conducted with a mixtureof Fmoc-protected amino acid (5 eq), HOBT (5 eq), HBTU (5 eq), DIEA (10eq) in DMF (0.1 mL), and NMP (0.2 mL) with agitation for 45 min. Thewashing cycle was repeated before the next round of deprotection andcoupling. After synthesis, peptides were washed in methanol and dried 24h. Protected peptide was cleaved with 1 mL of trifluoroacetic acid(TFA)/thioanisole/water/1,2-ethanedithiol (10/0.5/0.5/025) for 3 h atroom temperature and the resultant peptide solution was precipitated inmethyl tert-butyl ether.

Analytical and preparative HPLC was conducted, as described previously(5, 9), to purify each peptide to 80-90%. Correct peptide mass wasconfirmed by matrix-assisted laser desorption/ionization massspectroscopy (MALDI, Voyager instrument, Applied Biosystems), asdescribed previously (9). Measurements were made in linear, positive ionmode with an α-cyano-4-hydroxycinnamic acid matrix (data not shown).

Acetylated and benzoylated peptides derivatives (Ac135(L1)2_(—)1 andbz135(L1)2_(—)1, respectively) were synthesized by the same method usedfor N-terminal fluorescent labeling, described previously (7). Briefly,acetylation (acetic anhydrous) or benzyl addition (benzoatic acid) wasundertaken after assembly of the linear sequence by adding to the resin(10 eq in DIEA) for 2 h.

Binding of Targeting Peptides. The binding of peptides 1_(—)2, 1_(—)3,1_(—)4, 1_(—)6, 2_(—)1, and 3_(—)1 was assessed by fluorescentmicroscopy. S. mutans UA159 was grown overnight and diluted 1:5000 infresh TH with 1% sucrose before seeding to 96-well plates (flat bottom).Biofilms were grown 24 h and the spent medium replaced with 1×PBScontaining 25 μM peptide. After 5 min incubation at room temperature,the supernatants were removed and the biofilms washed 2× with 1×PBS andimaged by bright-field and fluorescence microscopy (Nikon E400). Digitalimages were collected and analyzed for semiquantitative bindingassessments with the manufacturers supplied software (SPOT, Diagnostics)(6).

Minimum Inhibitory Concentrations (MICs) of Peptides. Peptide MIC wasdetermined by broth microdilution (6, 15). Briefly, two-fold serialdilutions of each peptide were prepared with 50% BHI medium+50% sterilewater (for oral streptococci; all other bacteria were diluted in 1×Mueller-Hinton broth) at a volume of 100 μL per well in 96-wellflat-bottom microtiter plates. The concentration of peptide for thefirst test round ranged from 500 to 2 μg/mL or 125 to 1 μg/mL. Ifactivity was detected, a second round of MIC tests was conducted withconcentrations of 62.5 to 0.5 μg/mL. In either case, the microtiterplate was inoculated with a bacterial cell suspension to a finalconcentration of ˜1×10⁵ cells per mL and the plates were incubated at37° C. for 16-20 h under the appropriate conditions. After incubation,absorbance at 600 nm (A₆₀₀) was measured using a microplate UV-Visspectrophotometer (Model 3550, BioRad, Hercules, Calif.) to assess cellgrowth. The MIC endpoint was calculated as the lowest concentration ofantibacterial agent that completely inhibited growth or that produced atleast 90% reduction in turbidity when compared with that of apeptide-free control. At least 10 independent tests were conducted perpeptide. For peptides insoluble in aqueous solutions, stock solutionswere prepared in methanol or ethanol and appropriate solvent controlswere utilized. Cell growth was not affected by 5% (v/v) methanol orethanol, as described previously (10).

Peptide Killing Kinetics. To determine antimicrobial kinetics andspecificity, assays similar to traditional time-kill experiments wereperformed, as described previously (6, 7). Briefly, overnight bacterialcultures were diluted in BHI to A₆₀₀ 0.08 and peptides were added asindicated. Aliquots were then removed at various intervals and diluted1:50 in BHI and kept on ice until plating on appropriate growth medium.After 24 hours incubation, colonies were counted and the survivingcfu/mL determined. All assays were repeated at least 3 times and theaverage recovered cfu/mL were presented with standard deviations.Statistical analysis was conducted utilizing an unpaired Student'st-test.

Biofilm Growth Inhibitory Assays. STAMPs were tested for anti-biofilmactivity as described previously (6). Briefly, overnight cultures of S.mutans were diluted 1:50 in Todd-Hewitt (TH) broth medium supplementedwith 0.5% sucrose and 100 μL of bacterial suspension was added to eachwell of a 96-well microtiter plate. After centrifugation, bacteria werethen incubated under anaerobic conditions at 37° C. for 4 h.Supernatants were then removed and replaced with 25 μM peptide in 1×PBSfor 30 s to 1 min, followed by removal, washing, and replacement with100 μL fresh TH broth (without sucrose). Plates were then incubated at37° C. under anaerobic conditions and the bacterial recovery monitoredby recording A₆₀₀ after 4 h incubation. An unpaired Student's t-test wasutilized for statistical analysis.

Results

STAMPs consist of 3 regions: one targeting and one antimicrobial,connected via a flexible linker. In this report, we conjoined examplesof each domain with a variety of linkers to construct a pool of initialSTAMP candidates. These peptides were then evaluated for anti-S. mutansactivity and selectivity, their designs improved, and the lead STAMPsevaluated against S. mutans biofilms.

Selection of components and initial STAMP library design. As describedelsewhere, we generated several novel S. mutans-specific bindingpeptides, including 2_(—)4, (previously S3L1-10 (FIKDFIERF)) and 1_(—)5,(previously S3L1-5 (WWYNWWQDW)) (7). From these base sequences, residuesdiffering in hydrophobicity and/or charge were substituted at definedpositions to yield a series of related targeting sequences that werethen evaluated for binding to S. mutans biofilms. Several of the 1_(—)5variants, and one of the 2_(—)4 variants, were found to retain biofilmbinding (data not shown). These sequences (1_(—)2; 1_(—)3; 1_(—)4;1_(—)6; 2_(—)1; 3_(—)1), as well as 1_(—)5, were regarded in the presentstudy as the pool of S. mutans targeting peptides for STAMP construction(shown in Table 15). For the antimicrobial component, we selectedPL-135, a peptide based on an AMP isolated from tunicates (19), for theinitial round of design (Library 1). We hypothesized that linker regionsand attachment orientation would exert an influence on STAMP activity.Therefore, we initially conjugated each potential targeting peptide tothe N or C terminus of PL-135 through six different linkers, as shown inTable 15 (GGG, designated L1; SAT, L3; ASASA, L5; PYP, L7; PSGSP, L8;PSPSP, L9) leading to the synthesis of 84 STAMPs.

TABLE 15 STAMP constituent regions utilized in STAMP Library 1. S.mutans targeting Linker peptides peptides No. Sequence (name) Killingpeptide 1_2 WWHSWWSTW GGG (L1) FHFHLHF* (PL-135) 1_3 WWSYWWTQW SAT (L3)1_4 WWKDWWERW ASASA (L5) 1_5 WWYNWWQDW PYP (L7) 1_6 WWQDWWNEW PSGSP (L8)2_1 FIKHFIHRF PSPSP (L9) 3_1 LIKHILHRL *amidated C-terminus

Antimicrobial activity of initial STAMP library. To roughly gaugeLibrary 1 STAMP antimicrobial activity and S. mutans-selectivity, MIC(minimal inhibitory concentration) assays were conducted against S.mutans and a panel of bacteria, including two oral streptococci, S.sanguinis and S. sobrinus (FIG. 17). STAMPs containing 2_(—)1 conjoinedto the C-terminus of PL-135 (135(L1)2_(—)1, 135(L3)2_(—)1,135(L5)2_(—)1, 135(L7)2_(—)1, 135(L8)2_(—)1, 135(L9)2_(—)1), or 3_(—)1conjoined to the N-terminus of PL-135 (135(L1)3_(—)1) were found to beactive against S. mutans at concentrations lower than 100 μg/mL. Thesepeptides were more active (2-4 2-fold dilution steps) against S. mutansthan against the other oral streptococci or non-oral organisms tested.In contrast, native PL-135 had similar MICs against all strains examined(FIG. 17).

Impact of linker and terminal modification on STAMP antimicrobialactivity. We sought to investigate the impact of linker length or type,and N-terminal modification on the activity of STAMP 135(L1)2_(—)1 fromLibrary 1. As shown in Table 16, altered components included: 1) newlinkers GGGG (G₄) to GGGGGGG (G₇), AAA (L2), AGA (L10), GAGAG (L11),8-aminocaprylic acid (LC); 2) increased N-terminal aromacity (benzoicacid addition); 3) acetylation of N-terminus; or 4) component sequenceinversion (135i(L1)2_(—)1i). We observed MICs for these Library 2 STAMPsthat were similar to, or 2-fold less-active, than that of the base STAMP135(L1)2_(—)1 (FIG. 17), suggesting that the linker, termini andinversion alterations did not lead to improved activity against S.mutans, as measured by these means.

TABLE 16 Minimal inhibitory concentration of (MIC) of Library 2 STAMPsS. mutans No. Sequence^(a) (MIC)^(b) 135(L1)2_1 FHFHLHFGGGFIKHFIHRF 16135(G₄)2_1 FHFHLHFGGGGFIKHFIHRF 16 135(G₅)2_1 FHFHLHFGGGGGFIKHFIHRF 16135(G₆)2_1 FHFHLHFGGGGGGFIKHFIHRF 32 135(G₇)2_1 FHFHLHFGGGGGGGFIKHFIHRF32 135(L11)2_1 FHFHLHFSGSFIKHFIHRF 32 135(L12)2_1 FHFHLHFGSGSGFIKHFIHRF32 Ac135(L1)2_1 Ac-FHFHLHFGGGFIKHFIHRF 32 bz135(L1)2_1 benzoate- 32FHFHLHFGGGFIKHFIHRF 135(L2)2_1 FHFHLHFAAAFIKHFIHRF 16 135(L13)2_1FHFHLHFAGAFIKHFIHRF 32 135(L14)2_1 FHFHLHFGAGAGFIKHFIHRF 32 2_1i(L1)135iFRHIFHKIFGGGFHLHFHF 16 135i(L1)2_1i FHLHFHFGGGFRHIFHKIF 16 135(LC)2_1FHFHLHF-[NH(CH₂)₇CO]- 64 FIKHFIHRF ^(a)all C termini amidated ^(b)MIC(μg/mL) data from all S. mutans isolates tested, with a minimum of threeindependent trials.

Further potential of PL-135-based STAMPs. Antiseptic mouthrinses, suchas Listerine® (Warner-Lambert, Morris Plains, N.J.), are rapid-actingnon-selective bactericidal agents that can inactivate viable bacteriawithin seconds of contact (3). In order for STAMPs to be usefulmouthrinse ingredients, the antimicrobial kinetics must approach thisscale. Therefore, the killing kinetics of our Library 1 and 2 STAMPsfrom Table 15 and FIG. 17 were evaluated (data not shown). The resultsindicate that these PL-135-containing STAMPs, although selective for S.mutans when measured by MIC, are not rapid killers of this bacterium invitro, requiring several hours of exposure for observable antimicrobialactivity. Therefore, we sought to improve our STAMP pool by substitutingalternative killing domains for S. mutans STAMP construction.

Tuning the design of 2_(—)1-containing STAMPs. To improve theidentification of rapid-killing S. mutans STAMPs, we conjugated 2_(—)1with five AMPs selected from our previous studies (10), to constructLibrary 3: RWRWRWF (2c-4), FKKFWKWFRRF (B-33), IKQLLHFFQRF (B-38),RWRRLLKKLHHLLH (α-11), LQLLKQLLKLLKQF (a-7); attached at the C- orN-terminus. The linkers selected were L1, SGG (L2), L3 and LC. As shownin FIG. 18, MIC results indicate little difference in activity betweenconstructs where the targeting peptide was attached to the N or Cterminus of the AMP region, and little difference between linkersemployed. It was also apparent that these STAMPs were more activeagainst S. mutans relative to the other oral and non-oral bacteriatested: peptide 2_(—)1(L1)B33 demonstrated the lowest MIC range of 4 to8 μg/ml, which was a 2-4 fold improvement over the MIC for the killingpeptide alone (10). Taken together, these data suggest that Library 3STAMPs can effectively inhibit the growth of S. mutans atgenerally-improved potencies when compared to PL-135-containing STAMPsin Libraries 1 and 2.

STAMP killing kinetics against oral bacteria. Since the MIC assaymeasures antimicrobial activity after overnight incubation, largedifferences in killing rates between STAMPs and parental AMPs may beobscured in this assay, especially when the target organism issusceptible to the AMP (7, 9). To assess any significant selectivity andshort-term antimicrobial activity of the Library 3 STAMPs, time-killassays were performed against a variety of oral bacteria. Against thetargeted bacterium S. mutans (examples shown in FIG. 19D), the STAMPsacted significantly faster than the killing peptide alone within 5 minof treatment (p<0.001 comparing B-33 alone vs. 2_(—)1(L1)B33, or 2C-4vs. either 2C-4-containing STAMP). In contrast, other oral streptococci,such as S. mitis and S. gordonii, were less affected by STAMP treatments(FIG. 19A-C). Peptide 2_(—)1(L1)B33 exhibited the fastest killingkinetics and best selectivity: killing was observed even when cells weretreated for as little as 30 s, a timescale more appropriate for oralcavity therapeutic applications. As expected from their wide-spectra ofactivities (10), parental AMPs 2C-4 and B-33 had similar levels ofactivity against the strains examined.

Inhibition of biofilm growth. Although rapid and selective killing of S.mutans monocultures was apparent from the data shown in FIG. 19, wesought to determine whether these STAMPs would make suitableantimicrobial agents in the oral cavity, where multi-species biofilmsknown as dental plaque predominate (16, 18). To investigate, S. mutansbiofilms were treated with STAMPs and the post-antibiotic effect wasobserved after 4 h of biofilm recovery. As shown in FIG. 20, STAMPs2_(—)1(L1)2C-4, 2_(—)1(L6)2C-4, and 2_(—)1(L1)B33 were found tosignificantly inhibit (p<0.001) the formation of biofilms when cellswere treated with the peptide for 1 min at 25 μg/ml, compared tomock-treated biofilms or biofilms treated with untargeted AMP. Similarantimicrobial effects were observed for Listerine and Chlorhexidine.These results suggest that STAMP treatment persists after peptideremoval at a level similar to established wide-spectrum oralantiseptics.

DISCUSSION AND CONCLUSION

In this report, we present a novel strategy for the design and synthesisof STAMPs with activity against the oral pathogen S. mutans. Successfuldesign was achieved through a tunable, building-block approach thatutilized various combinations of antimicrobial, targeting, and linkerSTAMP peptide regions. Our results demonstrate that less-efficaciousSTAMPs could be improved when alternative killing regions weresubstituted in the design. While it remains unclear whether thealternative moieties were more active, or simply more conjugationtolerant, this process resulted in STAMPs that displayed killingkinetics against biofilms consistent with oral therapeutic applications.Additionally, more understanding was gained regarding AMP or targetingpeptide orientation dependencies, impact of linker regions, andappropriate targeting peptide choice, which could positively impactfuture anti-S. mutans STAMP design and refinement.

From the data presented here, it is difficult to determine the precisemechanism for the S. mutans-selectivity. Previous studies with STAMPshave indicated that the enhanced selectivity for the targeted strain isdue to the binding of the targeting peptide moiety (6, 7). Althoughdetailed binding analysis was not conducted in this study, our resultssuggest that a similar targeting-mediated killing is occurring here:targeting peptides, independently selected for STAMP construction on thebasis of their S. mutans-binding abilities, were required to enhance AMPantimicrobial activity and selectivity.

The data presented suggest that PL-135 may be inhibited by conjugationto other peptide subunits, as unmodified PL-135 displayed activityagainst S. mutans that was 2 to 4-fold higher than in progeny STAMPs, asshown in FIG. 17 and Table 16 (however, these constructs were selectivefor S. mutans). The unusually small size of this AMP may impart a severerestriction on amino acid additions, especially if the mode of actiondepends on sequence-dependent self-association on the target-cellmembrane, or binding to a discrete intracellular bacterial target (2).Our results suggest that AMPs with quicker “base” antimicrobial kinetics(such as B-33 or 2C-4), and higher tolerance for conjugations, should beselected for the design of STAMPs with optimal levels of enhancedkilling kinetics and selectivity.

Outside of PL-135-containing examples (and potentially some α-7 and α-11molecules, see FIG. 18), STAMPs in this report displayed no differencein activity between oppositely-oriented N and C-terminal AMP-targetingconjugations, suggesting that the optimal arrangement of STAMP domainsin likely AMP-specific, and depends on which least affects theantimicrobial mechanism. For example, the Pseudomonas spp-specificSTAMPs G10KHc and G10KHn (oriented target-killing and killing-target,respectively) both bind specifically to the target bacterium surface,but only G10KHc has significant membrane disruption activity (5, 7).

Interestingly, 2_(—)1 and 3_(—)1 containing STAMPs were active againstS. mutans, and the constructs with any other targeting peptide in Table15, were not. Targeting peptides 1_(—)2 through 1_(—)6 are stronglyhydrophobic, compared with 2_(—)1 and 3_(—)1 (8), and it may be possiblethat this characteristic limits the dissociation of these molecules fromthe hydrophobic components of the S. mutans cell wall, resulting intheir inhibitory affect on AMPs when conjugated, similarly to somestrong LPS-binding AMPs (14). However, the systematic building-blockdesign strategy employed allowed us to generate a diverse array ofSTAMPs, allowing us to identify useful compounds despite these stumblingblocks.

In conclusion, this report details the rational design of S.mutans-selective STAMPs with enhanced antimicrobial killing kinetics andselectivity when compared to untargeted AMPs. The S. mutans-selectiveSTAMPs were constructed using a tunable, combinatorial approach thatgenerated a diverse number of STAMP sequences for antimicrobialevaluation and improvement; a process may serve as an example for thesystematic development of novel selective antimicrobial agents. Wepropose that these STAMPs could be useful in the design of therapeuticsagainst oral or other mucosal pathogens, where the high diversity of“probiotic” beneficial microflora limits the effectiveness ofbroad-spectrum antimicrobial agents.

REFERENCES

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It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference in theirentirety for all purposes.

1. A chimeric moiety, said moiety comprising: an effector attached to apeptide targeting moiety comprising the amino acid sequence of a peptidefound in Table 2, Table 15, or any peptide disclosed herein.
 2. Thechimeric moiety of claim 1, wherein said targeting moiety is a peptideconsisting of the amino acid sequence of a peptide found in Table 2 orTable
 15. 3. The chimeric moiety of claim 1, wherein said effectorcomprises a moiety selected from the group consisting of a detectablelabel, an antimicrobial peptide, an antibiotic, and a photosensitizer.4. The chimeric moiety of claim 1, wherein said effector comprises anantimicrobial peptide comprising the amino acid sequence of a peptidefound in Table 2, Table 8, Table 9, or Table 10 or any antimicrobialpeptide disclosed herein.
 5. The chimeric moiety of claim 1, whereinsaid effector comprises an antibiotic found in Table
 7. 6. The chimericmoiety of claim 1, wherein said effector comprises a photosensitizer. 7.The chimeric moiety of claim 1, wherein said effector comprises aphotosensitizer selected from the group consisting of a porphyrinicmacrocycle, a porphyrin, a chlorine, a crown ether, an acridine, anazine, a phthalocyanine, a cyanine, a psoralen, and a perylenequinonoid.8. The composition of claim 7, wherein said photosensitizing agent is anagent shown in any of FIGS. 1-11.
 9. A chimeric moiety, said moietycomprising: a targeting moiety attached to an antimicrobial peptidewherein said antimicrobial peptide comprises the amino acid sequence ofa peptide found in Table 2 or any antimicrobial peptide disclosedherein.
 10. The chimeric moiety of claim 9, wherein said antimicrobialpeptide consists of the amino acid sequence of a peptide found in Table2.
 11. The chimeric moiety of claim 9, wherein said targeting moiety isa peptide comprising the amino acid sequence of a peptide found in Table2, Table 3, Table 4, Table 6, Table 15 or any peptide disclosed herein.12. The chimeric moiety of claim 9, wherein said targeting moiety is apeptide consisting of the amino acid sequence of a peptide found inTable 2 or Table
 15. 13. The chimeric moiety of claim 9, wherein saidtargeting moiety is an antibody that binds a microorganism.
 14. Thechimeric moiety of claim 13, wherein said antibody is an antibodyidentified in Table
 5. 15. The chimeric moiety according to any ofclaims 1-16, wherein said targeting moiety is chemically conjugated tosaid effector.
 16. The chimeric moiety of claim 15, wherein saidtargeting moiety is chemically conjugated to said effector via a linker.17. The chimeric moiety of claim 15, wherein said targeting moiety ischemically conjugated to said effector via a linker comprising apolyethylene glycol (PEG).
 18. The chimeric moiety of claim 15, whereinsaid targeting moiety is chemically conjugated to said effector via anon-peptide linker found in Table
 11. 19. The chimeric moiety accordingto any of claims 1-12, wherein said targeting moiety is linked directlyto said effector.
 20. The chimeric moiety according to any of claims1-12, wherein said targeting moiety is linked to said effector via apeptide linkage.
 21. The chimeric moiety of claim 20, wherein thechimeric moiety is a fusion protein.
 22. The chimeric moiety of claim21, wherein said linker is a peptide linker found in Table
 11. 23. Thechimeric moiety of claim 20, wherein said chimeric moiety isfunctionalized with a polymer to increase serum halflife.
 24. Thechimeric moiety of claim 21, wherein said polymer comprises polyethyleneglycol and/or a cellulose or modified cellulose.
 25. A pharmaceuticalcomposition comprising a chimeric moiety according to any of claim 1-24in a pharmaceutically acceptable carrier.
 26. The composition of claim25, wherein said composition is formulated as a unit dosage formulation.27. The composition of claim 25, wherein said composition is formulatedfor administration by a modality selected from the group consisting ofintraperitoneal administration, topical administration, oraladministration, inhalation administration, transdermal administration,subdermal depot administration, and rectal administration.
 28. Acomposition comprising an isolated antimicrobial moiety, said moietycomprising the amino acid sequence of a peptide found in Table
 2. 29.The composition of claim 28, wherein said peptide is a peptide selectedfrom the group consisting of a peptide consisting of the amino acidsequence of a peptide found in Table 2 comprising all L residues, apeptide consisting of the amino acid sequence of a peptide found inTable 2 comprising a peptide found in Table comprising all D residues, apeptide comprising the inverse of an amino acid sequence found in Table2, a peptide comprising the retro-inverso form of a peptide found inTable 2, a peptide found in Table 2 comprising a conservativesubstitution, and a peptide found in Table 2 comprising substitution ofa naturally occurring amino acid with a non-naturally occurring aminoacid.
 30. The composition of claims 29, wherein said peptide comprisesno more than 4 conservative substitutions.
 31. A composition comprisingan isolated targeting moiety, said moiety selected from the groupconsisting of a peptide comprising the amino acid sequence of a peptidefound in Table 2 comprising all L residues, a peptide consisting of theamino acid sequence of a peptide found in Table 2 comprising a peptidefound in Table comprising all D residues, a peptide comprising theinverse of an amino acid sequence found in Table 2, a peptide comprisingthe retro-inverso form of a peptide found in Table 2, a peptide found inTable 2 comprising a conservative substitution, and a peptide found inTable 2 comprising substitution of a naturally occurring amino acid witha non-naturally occurring amino acid.
 32. The composition of claim 31,wherein said peptide comprises no more than 4 conservativesubstitutions.
 33. A method of inhibiting the growth and/orproliferation of a microorganism and/or a biofilm comprising saidmicroorganism, said method comprising: contacting said microorganism orbiofilm with a composition comprising an antimicrobial peptidecomprising the amino acid sequence of a peptide selected from thepeptides in Table 2 or any antimicrobial peptide disclosed herein;and/or contacting said microorganism or biofilm with a compositioncomprising an antimicrobial moiety attached to a targeting peptidecomprising the amino acid sequence of a peptide selected from thetargeting peptides in Table 2, Table 15, or any targeting peptidedisclosed herein.
 34. The method of claim 33, wherein said microorganismor biofilm is a bacterium or a bacterial film.
 35. The method of claim33, wherein said targeting peptide is chemically conjugated to saidantimicrobial peptide.
 36. The method of claim 33, wherein saidtargeting peptide is linked directly to said antimicrobial peptide. 37.The method of claim 33, wherein said targeting peptide is linked to saidantimicrobial peptide via a linker comprising a polyethylene glycol. 38.The method of claim 33, wherein said targeting peptide is linked to saidantimicrobial peptide via a non-peptide linkage in Table
 11. 39. Themethod of claim 33, wherein said targeting peptide is linked to saidantimicrobial peptide via a peptide linkage.
 40. The method of claim 39,wherein said targeting peptide linked to said antimicrobial peptide is afusion protein.
 41. The method of claim 39, wherein said linker is apeptide linker in Table
 11. 42. A method of detecting a microorganism ora biofilm comprising said microorganism, said method comprising:contacting said microorganism or biofilm with a composition comprising adetectable label attached to a targeting peptide comprising the aminoacid sequence of a peptide selected from the peptides in Table 2, Table15, or any targeting peptide disclosed herein; and detecting saiddetectable label wherein the quantity and/or location of said detectablelabel is an indicator of the presence of said microorganism or biofilm.43. The method of claim 42, wherein said microorganism or a biofilm is abacterium or a bacterial film.
 44. The method of claim 42, wherein saidtargeting peptide consists of the amino acid sequence of a peptide foundin Table 2 or Table
 15. 45. The method of claim 42, wherein saiddetectable label is a label selected from the group consisting of aradioactive label, a radio-opaque label, a fluorescent dye, afluorescent protein, an enzymatic label, a colorimetric label, and aquantum dot.
 46. A composition comprising a photosensitizing agentattached to a targeting peptide comprising the amino acid sequence of apeptide found in Table
 2. 47. The composition of claim 46, wherein saidtargeting peptide consists of the amino acid sequence of a peptide foundin Table 2 or Table
 15. 48. The composition of claim 46, wherein saidphotosensitizing agent is an agent selected from the group consisting ofa porphyrinic macrocycle, a porphyrin, a chlorine, a crown ether, anacridine, an azine, a phthalocyanine, a cyanine, a psoralen, and aperylenequinonoid.
 49. The composition of claim 46, wherein saidphotosensitizing agent is an agent shown in any of FIGS. 1-11.
 50. Thecomposition of claim 46, wherein said photosensitizing agent is attachedto said targeting peptide by a non-peptide linker.
 51. The compositionof claim 46, wherein said photosensitizing agent is attached to saidtargeting peptide by a linker comprising a polyethylene glycol (PEG).52. The composition of claim 46, wherein said photosensitizing agent isattached to said targeting peptide by a non-peptide linker found inTable
 11. 53. A method of inhibiting the growth or proliferation of amicroorganism or a biofilm, said method comprising contacting saidmicroorganism or biofilm with a composition according to any of claims45-52.
 54. The method of claim 53, further comprising exposing saidmicroorganism or biofilm to a light source.
 55. The method of claim 53,wherein said microorganism is a microorganism selected from the groupconsisting of a bacterium, a yeast, a fungus, a protozoan, and a virus.56. The method of claim 53, wherein said biofilm comprises a bacterialfilm.
 57. The method of claim 53, wherein said biofilm is a biofilm onan implanted or implantable medical device.
 58. The method of claim 53,wherein said microorganism or biofilm is an organism or biofilm in anoral cavity.